THROMBOLYTIC (Blood Clot Destroyer)

MANDATORY FDA DISCLAIMER 

Introducing

THROMBOLYTIC

blood clot DESTROYER

200:1 Concentration

Featuring: Achyranthes Bidentata Bl. Root Extract • Aconitum Carmichaelii Debx. Root Extract • Acori Tatarinowii Rhizoma Extract • Albizia Richardiana Stem Extract • Alstonia Scholaris (L.) R. Br. Leaf Extract • Angelica Sinensis (Oliv.) Diels Root Extract • Anogeissus Latifolia Leaf Extract • Asparagi Radix Extract • Astragalus Membranaceus (Fisch.) Bge. Extract • Aucklandia Costus • Averrhoa Bilimbi (Oxalidiaceae) • Azadirachta Indica • Breviscapine • Bupleurum Chinense Dc. Extract • Carthamus Tinctorius FLOWer Extract • Chaenomeles Sinensis (Thouin) Koehne Extract • Cinnamomum Cassia Presl. Extract • Citrus Assamensis Leaf Extract • Citrus Aurantium L. Extract • Clerodendrum Viscosum (Verbanaceae) • Coicis Semen Extract • Cordycepin • Cordyceps Sinensis (Berk.) Sacc Extract • Curcumae Radix Extract • Cyathula Officinalis Kuan • Cyathulae Radix Extract • Dimocarpus Longan Lour. Extract • Dragon’s Blood From Dracaena Cochinchinensis (Lour.) S.C. Chen • Drynaria Quercifolia (Polypodiaceae). • Egb 761 From Ginkgo Biloba Extract • Epimeredi Indica (L.) Rothm • Evodia Rutaecarpa (Juss.) Benth Extract • Ganoderma Lucidum • Gastrodiae Rhizoma Extract • Haliotis Diversicolor Reeve Extract • Hawthorn Berry – Crataegus Laevigata • Kaempferol • Lagerstroemia Speciosa Leaf Extract • Leonurus Japonicus Sweet Extract • Licorice Root Extract -Glycyrrhiza Glabra L. • Ligusticum Chuam.Iong Hort.Extract • Lonicerae Japonicae Flos Extract • N-Acetylcysteine • Nattokinase • Nelumbinis Plumula Seed Extract • Nigella Sativa (L.) Seeds • Paeonia Suffruticosa Andr. Peel Extract • Paeonia Veitchii Lynch Root Extract • Panax Ginseng • Panax Notoginseng (Burk.)F. H. Chen Root Extract • Polygonum Multiflorum • Polygonum Tinctorium • Poria Cocos( Schw.) Wolf Extract • Pterocarpus Mildbraedii Harms Leaf Extract • Pueraria Lobata(Willd.)Ohwi Extract • Rehmanniae Radix Extract • Rutin • Salvia Miltiorrhiza Bge.Root Extract • Sargentodoxa Cuneata (Oliv.) Rehd. Et Wils. Extract • Sophora Japonical. Extract • Spirulan • Stevia Rebaudiana Leaf Extract • Taxillus Chinensis (Dc.) Danser.Extract • Tetracera Sarmentosa (L.) Vahl • Typha Angusti Folial Extract • Vitamin B6 • White Willow (Salix Alba)

Covid-19 blood clots

vaccine blood clotS

Sudden Death FROM BLOOD CLOTS

LONG Covid & Blood Clots

Myocarditis

pericarditis

MYOCARDIAL INFARCTION

POLYPHENOLS TO TREAT & PREVENT COVID-19

NATURAL ANTICOAGULANTS

PREVENTING BLOOD CLOTS WITH FLAVONOIDS & POLYPHENOLS

MEDICINAL PLANT THROMBOLYTICS

Anti-thrombotic activity of plant extracts

INGREDIENTS & SCIENTIFIC STUDIES:

Achyranthes bidentata Bl. root extract

Achyranthes bidentata polypeptide k improves long-term neurological outcomes through reducing downstream microvascular Thrombosis in experimental Ischemic Diseases Stroke

Achyranthes bidentata Bl. (A. bidentata) occupies an important position in traditional Chinese medicine owing to the property of promoting the circulation of blood and removing stasis. Achyranthes bidentata polypeptide k (ABPPk) is one of the active components isolated from A. bidentata. We previously demonstrated that ABPPk has potent neuroprotective effects against neuronal apoptosis both in vitro and in vivo, but the roles and mechanisms of ABPPk on long-term functional recovery after Ischemic Diseases Stroke remain unknown. In the current study, we investigated the neuroprotective effects of ABPPk on filament transient middle cerebral Artery occlusion (tMCAO) rats and found that ABPPk reduced the infarct volume and maintained the neuronal integrity in the Ischemic Diseases penumbra.

Moreover, we found that ABPPk might reduce the formation of downstream microthrombus through preventing Ischemic Diseases -induced oxidative damage of brain endothelial cells and activation of tissue factor (TF), plasminogen activator inhibitor-1 (PAI-1), and NF-κB. ABPPk also inhibited polymorphonuclear leukocytes (PMNs) infiltration and matrix metalloproteinase-2/-9 (MMP-2/-9) activation in the Ischemic Diseases penumbra. Morris water maze, foot fault test, and modified neurological severity score were assessed for a period of 6 weeks following tMCAO. ABPPk improved long-term recognition abilities and neurological outcomes after Stroke compared with saline-treated rats. Taken together, these results suggested that ABPPk is beneficial to the improvement of long-term outcomes after transient cerebral ischemia injury and can be used as a potential neuroprotective agent .

Aconitum carmichaelii Debx. root extract

A review on phytochemistry and pharmacological activities of the processed lateral root of Aconitum carmichaelii Debeaux

Aim of the review: The present paper aims to provide an up-to-date review at the advancements of the investigations on the ethnopharmacology, phytochemistry, pharmacological effect and toxicity of Fuzi. Besides, the possible tendency and perspective for future research of this plant are discussed, as well.

Materials and methods: All available information on Fuzi was collected via electronic search (using Elsevier, PubMed, ACS, CNKI, Google Scholar, Baidu Scholar, and Web of Science), books and classic works about Chinese herb.

Acori Tatarinowii Rhizoma extract

Evaluation of the effects of Acorus gramineus and Acorus tatarinowii Extracts on a Rat Model of Arterial Thrombosis induced by Ferric Chloride

We investigated the antithrombotic effect of Acorus gramineus Soland (A. gramineus) from Korea and Acorus tatarinowii Schott (A. tatarinowii) from China in a rat model of Arterial Thrombosis induced by ferric chloride ($FeC{l}_3$). Thirty minutes prior to a 35% $FeC{l}_3$ application, Sprague Dawley rats were intraperitoneally injected with saline, A. gramineus and A. tatarinowii (100 mg/kg), respectively. Occlusion time of rats injected with A. gramineus was delayed significantly compared to that of the vehicle and A. tatarinowii. thrombus weight was meaningfully decreased in rats injected with A. gramineus compared to the vehicle.

Additionally, A. gramineus inhibited collagen fiber damage in vessel compared to the vehicle, but A. tatarinowii did not show a significant effect. Our results show that A. gramineus and A. tatarinowii from the same genus have different antithrombotic effects , and especially A. gramineus has a better antithrombotic effect than A. tatarinowii.

effects of Acori Rhizoma Extract on the in vitro anti-platelet Activity in Human Whole blood

Objectives: Acori Rhizoma is one of the common widely used herbal medicines with diverse bioactive effects . However, little evidence has been reported about the potential anti-platelet activity of Acori Rhizoma. The present study examined the effects on platelet aggregation by Acori Rhizoma.

Methods: In this study, we tested the in vitro effect of 16 kinds of Acori Rhizoma extracts by hot water or 70% ethanol on collagen-induced platelet aggregation in human whole blood using the impedance method of aggregometry.

Results: Among them, 2 kinds of 70% ethanol extract and 1 kind of hot water extract showed the significant inhibiting effect on whole blood aggregation . In particular, Acorus gramineus extracts were selected as the most effective candidate.

Conclusions: The results from this experiment provide pharmacological evidence for the traditional medicine, suggesting that Acorus gramineus could be help problems of blood circulation more than Acorus tatarinowii.

Alstonia scholaris

In vitro Study of Different Partitionates of Alstonia scholaris (L.) R.Br. Leaf for Thrombolytic and Membrane Stabilizing Activities

Alstonia scholaris (L.) R.Br. is a medicinal herb belonging to the family Apocyanaceae. Locally it is known as Chatim and abundant with ethno-medicinal properties . The crude ethanolic extract of A. scholaris leaves along with its n-hexane, ethyl acetate, dichloromethane and aqueous soluble partitionates were assayed for their probable Thrombolytic and membrane stabilizing activities.

The activities of the leaf extract were compared to standard drugs, streptokinase for Thrombolytic and acetyl salicylic acid for membrane stabilizing activities. In this study, among all extractives the highest Thrombolytic activity was exhibited by the crude ethanolic extract (55.33±0.08%) (p < 0.001) as compared to Streptokinase (62.07±0.33%) (p < 0.001).

Alternatively, ethanol soluble materials also significantly inhibited the haemolysis of human erythrocyte membrane both in induced by hypotonic solution (83.47±0.15%) (p < 0.001) and by heat (79.82±0.71%) (p < 0.001), respectively as compared to standard acetyl salicylic acid (85.09±0.8%) (p < 0.001) and (80.98±1.34%) (p < 0.001).

Mechanisms underlying the antihypertensive effect of Alstonia scholaris

Ethnopharmacological relevance: Alstonia scholaris has a long history of use in the Ayurveda traditional treatment of various ailments including hypertension . We have reported the blood pressure lowering activity of the extract of A. scholaris. The following research aim to delineate the pharmacological mechanism involve in the antihypertensive action.

Materials and method: Vasorelaxant effect of the n-butanol fraction of A. scholaris (NBF-ASME) was evaluated on rat aorta pre-contracted with phenyelphrine (PE, 1 µM). Aortic rings preparation were pre-incubated with various antagonists like 1H-[1,2,4] oxadiazolo-[4,3-a]quinoxalin-1-one (ODQ 10 μM), methylene blue (MB 10 μM), Nω-nitro-L-arginine methyl ester hydrochloride (l-NAME 10 μM), atropine (10 μM), indomethacin (1 μM), ML-9 and various K+ channel blockers such as glibenclamide (10 μM) and tetraethyl ammonium (TEA 10 μM) for mechanism study.

Results: The results showed that pre-incubation of aortic rings with the extract (0.5, 1 and 2 mg/mL) significantly inhibit the contractile response of the rings to phenylephrine-induced contraction (p<0.05–0.001). Removal of endothelium, incubation with l-NAME, indomethacin, atropine and propranolol did not significantly affect the relaxation effect of NBF-ASME. Furthermore, the K+ channel blockers, TEA and glibenclamide showed no inhibitory effect. However, aortic rings pretreated with ODQ and ML-9 showed a significant suppression of the relaxation curve of NBF-ASME (p<0.01–0.001). In Ca2+-free solution, NBF-ASME inhibits the release of intracellular Ca2+ from the sarcoplasmic reticulum. NBF-ASME also inhibits calcium chloride (CaCl2)-induced contraction in endothelium-denuded aortic rings.

Conclusion: The results from this study suggests that A. scholaris exerts vasodilation via calcium channels blockade, direct activation of soluble guanylate cyclase and possibly by also inhibiting the formation of inositol 1, 4, 5-triphosphate.

Angelica sinensis (Oliv.) Diels root extract

Z-ligustilide Extracted from Radix Angelica Sinensis Decreased platelet aggregation Induced by ADP Ex Vivo and Arterio-Venous Shunt Thrombosis In Vivo in Rats

antithrombotic therapy has become an important goal for the treatment of Ischemic Diseases disorders such as cerebral ischemia . Our recent studies found that Z-ligustilide (LIG), a characterized 3-n-alkylphthalide constituent of Radix Angelica sinensis essential oil, exerted significant neuroprotection against cerebral Ischemic Diseases damage in several animal models. The present study evaluated the antithrombotic activity of LIG and its effect on platelet aggregation and Coagulation time. LIG (10 or 40 mg/kg) was intragastrically administered to rats once daily for 3 days. Our results showed that LIG significantly and dose-dependently reduced Arterial thrombus weight in an arterioVenous shunt Thrombosis in rats and platelet aggregation induced by adenosine diphosphate in rats ex vivo. Meanwhile, LIG at 10 or 40 mg/kg had no significant effect on Coagulation time, including activated partial thromboplastin time and prothrombin time, in rats ex vivo. The present study demonstrated for the first time that LIG may exert efficient antithrombotic activity through inhibition of platelet aggregation , without effecting Coagulation time of peripheral blood. These data, together with the previously reported neuroprotective effects of LIG on cerebral ischemia , suggest that the antithrombotic activity of LIG may contribute to its potential for the treatment of Ischemic Diseases Diseases , including Ischemic Diseases Stroke .

Polysaccharides from Angelica sinensis alleviate neuronal cell injury caused by oxidative stress

Angelica sinensis has antioxidative and neuroprotective effects . In the present study, we aimed to determine the neuroprotective effect of polysaccharides isolated from Angelica sinensis. In a preliminary experiment, Angelica sinensis polysaccharides not only protected PC12 neuronal cells from H₂O₂-induced cytotoxicity, but also reduced apoptosis and intracellular reactive oxygen species levels, and increased the mitochondrial membrane potential induced by H₂O₂ treatment. In a rat model of local cerebral ischemia , we further demonstrated that Angelica sinensis polysaccharides enhanced the antioxidant activity in cerebral cortical neurons, increased the number of microvessels, and improved blood flow after ischemia . Our findings highlight the protective role of polysaccharides isolated from Angelica sinensis against nerve cell injury and impairment caused by oxidative stress.

Medicinal plants with antithrombotic property in Persian medicine: a mechanistic review

Thrombosis is one of the major causes of morbidity and mortality in a wide range of vessels Diseases . Due to the high prevalence of Thromboembolic disorders investigations are being carried out on new antithrombotic agents with limited adverse side effects in which herbal medicines are considered as alternative remedies. Persian medicine (PM) as a traditional medicine has a good potential for pharmacotherapy based on its own principles and development of drugs via investigating PM literature. In PM manuscripts there are some concepts that express the management of blood Clots and antithrombotic properties.

This study reviewed the pharmacological effects of medicinal plants mentioned in PM literature for blood Clot management in light of current knowledge. Plants mentioned in PM for management of blood Clot belong to 12 families in which Apiaceae, Lamiaceae and Compositae were the most repeated ones. Among the proposed plants Allium sativum, Rosmarinus officinalis, Boswellia serrata, Sesamum indicum, Matricaria chamomilla and Carthamus tinctorius have been the most researched plants in modern antithrombotic studies while for some plants such as Helichrysum stoechas, Dracocephalum kotschi, Carum carvi, Bunium persicum and Lagoecia cuminoides no evidence could be found. One of the interesting notes in Clot management in PM texts was introducing the target organ for some of the recommended herbs like Carum carvi and Bunium persicum for dissolving blood Clot in stomach and Commiphora mukul for thrombosed hemorrhoid.

It seems review of PM recommendations can help to design future researches for antithrombotic drugs discovering with more effectiveness and safety.

Anti-coagulant activity of plants: mini review

Medicinal plants have been used for treatment of human ailments since ancient times. Objective of this study is to document the effect of herbal drugs on Anticoagulant therapy. The material for this review was taken mostly from PubMed and the Cochrane database of systematic reviews. Some other relevant references were collected from personal database of papers on anti-coagulant properties of plants.

Literature review shows that many plants such as Thymus vulgaris, Cyamopsis tetragonoloba taub, Pulmonaria officinalis and Cinnamomum cassia etc have anti-coagulant activity. This review shows that medicinal plants should be prescribed with care to patients on Anticoagulant therapy.

Stroke therapy in traditional Chinese medicine (TCM): prospects for drug discovery and development

Brain injuries resulting from Stroke are a major and increasing public health problem in both developed and developing countries worldwide. China’s extensive experience in the use of traditional Chinese medicines (TCMs) in Stroke therapy indicates that TCM preparations are effective, with few or no side-effects . There are more than 100 traditional medicines in use for Stroke therapy in China. Some of their therapeutic effects in Stroke have been confirmed by recent clinical studies. A large number of compounds have been isolated from TCMs and most of these resources have not yet been characterized for pharmacological purposes.

Here, this article explains how TCM provides an extensive and knowledge-rich foundation for implementing a strategically focused pharmacological research programme aimed at the development of new drugs.

Overview of therapeutic potentiality of Angelica sinensis for Ischemic Diseases Stroke

Background: Ischemic Diseases Stroke is a common cerebrovascular Disease . Due to sudden interruption of blood flow by Arterial thrombus, amounts of neurons in Ischemic Diseases central and penumbral regions occur necrosis and apoptosis resulting in serious injury of neurological function. Chinese medicines have a great advantage in Ischemic Diseases Stroke treatment and recovery, especially Angelica sinensis.

Purpose: There are a large number of studies reported that Angelica injection and A. sinensis active compounds. We systematically reviewed the effects and mechanisms of A. sinensis in recent years according to the guidelines of the Preferred Reporting Items for Systematic Review and Meta-Analysis (PRISMA) statements, and excavated its therapeutic potentiality for exploring more effective and safe compounds for Ischemic Diseases Stroke precision treatment.

Results: A. sinensis extracts and active compounds, such as Z-ligustilide, 3-n-Butylphthalide, and ferulic acid have significant effects of anti-inflammation, anti-oxidative stress, angiogenesis, neurogenesis, anti-platelet aggregation, anti-atherosclerosis, protection of vessels, which contributes to improvement of neurological function on Ischemic Diseases Stroke.

Conclusion: A. sinensis is a key agent for Ischemic Diseases Stroke treatment, and worth Deeply excavating its therapeutic potentiality with the aid of pharmacological network, computer-aided drug design, artificial intelligence, big data and multi-scale modelling techniques.

Albizia richardiana

Stem extract of Albizia richardiana exhibits potent antioxidant , cytotoxic, antimicrobial, anti-inflammatory and Thrombolytic effects through in vitro approach

Anogeissus latifolia leaf extract

IN-VITRO Thrombolytic POTENTIAL OF LEAF EXTRACT OF
ANOGEISSUS LATIFOLIA

Thrombotic disorders such as Myocardial Infarction and cerebral impediment are fatal blood Clotting related Diseases . The need of safer and reliable herbal medicines for targeted and better drugs is the need of the time, since the Thrombosis may further lead to complex blood dyscrasias. Thus the current study is targeted at exploration of the Thrombolytic potential of the hydro alcoholic leaf extract of indigenous herb Anogeissus latifolia. The plant has shown presence of phytoconstituents such as tannins, polyphenols which has been reported to show good cardio protective activity. Also, the plant has reported to show antihyperlipidemic activity, good anti-oxidant and anti-atherosclerosis activity.

Hence the plant promises to show fair Thrombolytic potential. Streptokinase was used as the standard drug for comparison and the saline solution as vehicle control. The plant has shown significant activity compared to both the groups.

Asparagi Radix extract

Anti-Thrombic activity of Korean herbal medicine, Dae-Jo-Whan and its herbs

The anti-Thrombic properties of the Korean herbal medicine, Dae-Jo-Hwan (DJW) were investigated. Water extracts, a 70% methanol (MeOH) extract and an ethyl acetate (EtOAc) soluble fraction (III) from DJW inhibited platelet -activating factor (PAF)-induced platelet aggregation in vitro and in vivo assays. The extracts of DJW and eleven herbs from which it is derived, except for Panax ginseng Meyer, Angelica sinensis (OLIV.) DIELS and Schisandra chinensis Baill, inhibited AA-induced blood platelet aggregation to various extents.

The effects observed with total DJW was synergistic over-additive rather that additive since the sum of single contributions was lower than the effect of the total extract. Fraction III was specially protected against the lethality of PAF, while verapamil did not afford any protection. Exogenously applied arachidonic acid (AA) (100 μM) led to a 89% platelet aggregatio , the release of 14 pmol of ATP, and the formation of either 225 pg of thromboxane A2 (TXA2) or 45 pg of prostaglandin E2 (PGE2), each parameter being related to 106 platelet s. An application of DJW 5 min before AA, dose-dependently diminished aggregation, ATP-re lease, and the synthesis of TXA2 and PGE2, with IC50 values of 70, 87, 65 and 72 μg/ml, respectively.

The similarity of the IC50 values suggests the inhibition of cyclooxygenase (COX) by DJW as the primary target, thus suppressing the generation of TXA2, which induces platelet aggregation and the exocytosis of ATP by its binding on TXA2-receptors. These results indicate that DJW shows anti-thrombotic action on human platelets and inhibits the action of PAF in vivo by an antagonistic effect on PAF. Therefore, it may be useful in treating disorders caused by PAF.

Astragalus membranaceus(Fisch.)Bge.extract

The combination of Astragalus membranaceus extract and ligustrazine to improve the inflammation in rats with Thrombolytic cerebral ischemia

The purpose of the study was to evaluate the effect of Astragalus membranaceus extract and ligustrazine combination on ameliorating inflammation in cerebral Ischemic Diseases rats that have undergone thrombolysis . Astragalus membranaceus and ligustrazine per se, or a combination of A. membranaceus and ligustrazine was administered by intraperitoneal injection immediately after surgery and sham surgery. After the induction of thrombolysis, the neurological function was measured and cerebral lesion volume was determined. The regulatory T cells in the spleen were measured by flow cytometry. To explore the protective effects of the combination drug on the neurological function and inflammation, the expression of transcription factor Foxp3 and cytokines, including transforming growth factor beta 1, interleukin 10, interleukin 4, interleukin 1 beta, interferon gamma, interleukin 17, in damaged brain was examined using reverse transcription polymerase chain reaction,

Western blot, and enzyme-linked immunosorbent assay. The cerebral lesion volume was markedly reduced in the combination drug–treated rats compared to the rats treated with either A. membranaceus or ligustrazine alone (P < 0.05). The neurological function, regulatory T cells, transcription factor Foxp3, transforming growth factor beta 1, interleukin 10, and interleukin 4 were markedly elevated in the rats treated with combination drugs (P < 0.05). The expression of interleukin 1 beta, interferon gamma, and interleukin 17 was reduced in the rats treated with combination drug therapy (P < 0.05).

Treatment with a combination of A. membranaceus and ligustrazine can ameliorate inflammation after thrombolysis and regulate the related cytokines by elevating the expression of endogenous regulatory T cells.

The Combination of Astragalus membranaceus and Ligustrazine Protects Against thrombolysis -Induced Hemorrhagic Transformation Through PKCδ/Marcks Pathway in Cerebral ischemia Rats

Astragalus membranaceus (Ast) and ligustrazine (Lig) have a protective effect on lower hemorrhagic transformation induced by pharmaceutical thrombolysis . The cerebral ischemia rat model was induced with autologous blood Clot injections. A combination of Ast and Lig, or a protein kinase C delta (PKCδ) inhibitor—rottlerin, or a combination of Ast, Lig, and rottlerin was administered immediately after recombinant tissue plasminogen activator injection. The cerebral infarct area, neurological deficits, cerebral hemorrhage status, neuronal damage and tight junctions’ changes in cerebral vessels, and the messenger RNA and protein levels of PKCδ, myristoylated alanine-rich C kinase substrate (Marcks), and matrix metallopeptidase 9 (MMP9) were determined after 3 h and 24 h of thrombolysis.

The ultrastructure of the neuronal damage and tight junctions was examined under a transmission electron microscope. The expression levels of PKCδ, Marcks, and MMP9 were assessed by immunohistochemistry, western blot, and quantitative real-time polymerase chain reaction . Administration of Ast and Lig not only significantly decreased neurological deficit scores, infarct volumes, and cerebral hemorrhage but also inhibited the disruption due to neuronal dysfunction and the tight junction integrity in the cerebral vessel.

Treatment with a combination of Ast and Lig effectively protected ischemia-induced microhemorrhage transformation through PKCδ/Marcks pathway suppression.

The combination of Astragalus membranaceus extract and ligustrazine to improve the inflammation in rats with Thrombolytic cerebral ischemia c

The purpose of the study was to evaluate the effect of Astragalus membranaceus extract and ligustrazine combination on ameliorating inflammation in cerebral Ischemic Diseases rats that have undergone thrombolysis. Astragalus membranaceus and ligustrazine per se, or a combination of A. membranaceus and ligustrazine was administered by intraperitoneal injection immediately after surgery and sham surgery.

After the induction of thrombolysis, the neurological function was measured and cerebral lesion volume was determined. The regulatory T cells in the spleen were measured by flow cytometry. To explore the protective effects of the combination drug on the neurological function and inflammation, the expression of transcription factor Foxp3 and cytokines, including transforming growth factor beta 1, interleukin 10, interleukin 4, interleukin 1 beta, interferon gamma, interleukin 17, in damaged brain was examined using reverse transcription polymerase chain reaction, Western blot, and enzyme-linked immunosorbent assay.

The cerebral lesion volume was markedly reduced in the combination drug–treated rats compared to the rats treated with either A. membranaceus or ligustrazine alone (P < 0.05). The neurological function, regulatory T cells, transcription factor Foxp3, transforming growth factor beta 1, interleukin 10, and interleukin 4 were markedly elevated in the rats treated with combination drugs (P < 0.05). The expression of interleukin 1 beta, interferon gamma, and interleukin 17 was reduced in the rats treated with combination drug therapy (P < 0.05). Treatment with a combination of A. membranaceus and ligustrazine can ameliorate inflammation after thrombolysis and regulate the related cytokines by elevating the expression of endogenous regulatory T cells.

Astragalus Membranaceus and Ligustrazine Combination Ameliorates Micro-hemorrhage Through Maintaining blood-brain Barrier Integrity in Cerebral Ischemic Diseases Rats

Ethnopharmacological relevance: Hemorrhagic transformation is a frequent asymptomatic event that occurs after acute Ischemic Diseases Stroke, especially after pharmaceutical thrombolysis. However,the mechanism of hemorrhagic transformation and therapeutic s were unknown. In this study, we combine administered astragalus membranaceus and ligustrazine in rats underwent cerebral ischemia with thrombolysis; and analyzed their effects on attenuation of hemorrhagic transformation,maintaining of blood-brain barrier integrity.

Methods: A rat model of focal cerebral ischemia was induced with autologous blood Clot injection. Thrombolysis was performed via intraVenous injection of rt-PA.Astragalus membranaceus,ligustrazine or the combination of both were administered immediately after Clot injection. Cerebral infarct area,neurological deficits, blood-brain barrier integrity,and cerebral hemorrhage were determined after 3,6 and 24 hours of ischemia. Uitrastructure of blood-brain barrier was analyzed using transmission electron microscope. Tight junction protein including claudin-1,claudin-5,occludin,zonula occludens-1 expression and matrix metallopeptidase-9activation were further evaluated the role of combination drug on the protection of blood-brain barrier integrity.

Results: ischemia induced Evens blue leakage and cerebral hemorrhage were sufficiently reduced in combination treated rats compared to the astragalus membranaceus or ligustrazine alone(p0.05).The disruption of blood-brain barrier uitrastructure and neurological deficits were ameliorated after combination treatment(p0.05).It is noted that claudin-1,claudin-5,occludin and ZO-1 expression were less decreased in rats with combination treatment.In addition,MMP-9 activity was suppressed in the combination treated rats compared to the control(p0.05).

Conclusion: Combination treatment with astragalus membranaceus and ligustrazine alleviate ischemia-induced micro-hemorrhage transformation,which is through maintaining blood-brain barrier integrity.

Aucklandia costus

Aucklandia costus (Syn. Saussurea costus): Ethnopharmacology of an endangered medicinal plant of the himalayan region

Ethnopharmacological relevance: Aucklandia costus Falc. a medicinal plant is native to the Himalayan region and synonymous with Saussurea costus, Saussurea lappa, and Aucklandia lappa. It has an ancient background of being used ethnopharmacologically for various body ailments. According to Ayurveda, Unani, Siddha, and Traditional Chinese Medicine, Costus roots are recommended for leukoderma, liver, kidney, blood disorders , Qi stagnation, and tridosha. Root and powder are used orally with warm water to cure gastric problems, and the paste is applied to the inflamed area to relieve pain. Root paste is applied on the skin to cure boils, blisters, and leprosy.

Aim of the study: The aim of the present review is to establish a correlation among the ethnopharmacological uses and scientific studies conducted on A. costus with chemical constituents, safety & toxicity data including future directions for its conservation with higher yield and effect.

Materials and methods: The study was conducted by studying books, research papers, and literature in history, agroforestry, phytopharmacology of Himalayan plants using international databases, publication, Red data book, and reports. The search engines: Pubmed, Scopus, Wiley Inter-science, Indian Materia Medica, Science Direct, and referred journals are referenced.

Results: The literature collected from databases, journals, websites, and books mentioned the use of costus roots in local and traditional practices. CITES included A. costus in a critically endangered category due to lack of cultural practices and overexploitation from wild. A. costus roots are known since 13th century for use in ancient Ayurvedic products but the scientific evaluation is of future research interest. A correlation of traditional uses with scientific studies has been explored to assess the effect of root powder, extract, oil and isolated constituents: Costunolids, Saussureamine B and Dehydrocostus lactone etc. in gastric ulceration and lesions; inhibition of antigen-induced degranulation, mucin production, number of immune cells, eosinophils, and expression and secretion of Th2 cytokines (IL-4 and IL-13) in asthma. The inhibition of pro-inflammatory mediators is also reported by Cynaropicrin, Alantolactone, Caryophyllene, Costic acid. Also, the sesquiterpene lactones has profound effect in inhibition of inflammatory stages and induced apoptotic cascades in cancer. Very few data on the safety and toxicity of plant parts have been noted which needs to be evaluated scientifically.

Conclusion: A. costus have been noted to have remarkable effect for gastric, hepatic, inflammatory, respiratory, cancer, skin problems but there were several errors in selection of plant material, authentification, selection of dose, assessment, selection of standard and control have been identified. Therefore, a schematic drug development and research strategy exploiting the potential of plant extract, fraction, products and probable constituents, costunolide, dehydrocostus lactone, cynaropicrin, saussureamine assuring dose-response relationship and safety may be determined under pre-clinical which may be extrapolated to clinical level. An evaluation of phytochemicals in A. costus collected from different geographical location in Himalayas may be drawn to identify and conserve the higher yielding plant.

Averrhoa bilimbi

Evaluation of Thrombolytic potential of three medicinal plants available in Bangladesh, as a potent source of Thrombolytic compounds

A review on phytochemical constituents and biological assays of Averrhoa bilimbi,

Averrhoa bilimbi is a multipurpose, long-lived tropical plant commonly known as-Bilimbi, Cucumber Tree‖ belonging to family Oxalidaceae. The plant has an enormous fiscal value since most of the parts like leaves, bark, flow ers, fruits, seeds, roots or the whole plant are used as alternative medicine to treat a variety of Diseases especially diabetes.

In the present review, we tried to give the existing information on photochemical constituents, conventional medicinal uses and anti-microbial, anti-inflammatory , cyto-toxic activities, anti-oxidant activity, antifertility, and antibacterial activities and other biological activities of Averrhoa bilimbi.

The extract of various parts of Averrhoa bilimbi is medicinally used as a folk remedy for many symptoms and showed significant pharmacological activities so it is necessary to perform further investigation to isolate such pharmacological active compounds which can be used in production of novel drugs for various Diseases. Overall, this paper gives an overview on covering the biology, and various commercial and therapeutic applications.

Evaluation of fibrinolytic efficacy of Averrhoa Bilimbi linn. by using euglobulin lysis time method

Background: An impaired fibrinolytic function had been reported in subjects suffering from Venous Thrombosis . The euglobulin Clotlysis time is a test that reflects the overall fibrinolytic activity of plasma.

Objective: To evaluate the fibrinolytic efficacy of ethanolic extract of Averrhoa bilimbi Linn. leaves by using Euglobulin lysis time method.

Methods: Rats were divided into 3 groups. Each group was containing 6 rats. Group I: received vehicle (control) 0.9% saline solution; Group II: received 25 mg/kg ethanolic extract leaves of Averrhoa bilimbi Linn; Group III: received 50 mg/kg ethanolic extract leaves of Averrhoa bilimbi Linn. blood (1.8 ml) was removed with a plastic syringe containing 0.2 ml 3.8% sodium citrate solution. The time interval between the addition of thrombin and the complete lysis of the Clot s was measured. The lysis time (min) was determined.

Conclusion: We observed ethanolic extract of Averrhoa bilimbi Linn. leaves may increase the activators of the fibrinolysis. Further investigation is warranted.

Phytochemical screenings, Thrombolytic activity and antimicrobial properties of the bark extracts of Averrhoa bilimbi

In this present study, the bark extracts of Averrhoa bilimbi were subjected to the Thrombolytic activities were assessed by using human erythrocyte and the results were compared with standard streptokinase (SK). On the other hand, bark extracts of A. bilimbi revealed moderate antibacterial activity against some microorganisms used in the screening. Preliminary phytochemical investigation suggested the presence of flavonoids, saponins and alkaloids.

Azadirachta indica

Molecular docking and ADME/T analysis for identification of novel potential COX inhibitors of some isolated compounds from Azadirachta indica for Thrombolytic treatment

Developing a new agent in the Thrombolytic field, plants are widely used for the management of Myocardial Infarction , Thromboembolic Strokes, Deep vein Thrombosis and Pulmonary Embolism . For this purpose we subjected the active compounds of to reveal its potentiality by molecular docking analysis to find out its potent compound against 1A5H which was done by Maestro v 10.1 (Schrodinger) docking analysis. Docking studies by Maestro v 10.1 (Schrodinger) showed that Gamma-Elemene of Azadirachta indica had the lowest docking score respectively against the 1A5H which are-3.001. Gamma-Elemene from Azadirachta indica detected with significant docking score which may be a potent Thrombolytic compound because the less docking score, the compound will be more potent.

Thrombolytic potential of Ocimum sanctum L., Curcuma longa L., Azadirachta indica L. and Anacardium occidentale L.

Atherothrombotic Diseases such as Myocardial or cerebral Infarction are serious consequences of the thrombus formed xin blood vessels. Thrombolytic agent s are used to dissolve the already formed Clots in the blood vessels; however, these drugs have certain limitations which cause serious and sometimes fatal consequences. Herbal preparations have been used since ancient times for the treatment of several Diseases . The aim of this study was to investigate whether herbal preparations possess Thrombolytic activity or not. An in vitro Thrombolytic model was used to check the Clotlysis effect of four aqueous herbal extracts viz., O. sanctum, C. longa, A. indica, A. occidentale along with Streptokinase as a positive control and water as a negative control. The percentage (%) Clotlysis was statistically significant (p<0.0001) when compared with vehicle control.

Using an in vitro Thrombolytic model, O. sanctum, C. longa, A. indica & A. occidentale showed moderate Clotlysis activity (30.01 ± 6.168%, 32.94 ± 3.663%, 27.47 ± 6.943%, 33.79 ± 2.926% respectively) whereas standard streptokinase showed 86.2 ± 10.7 % Clotlysis effect. From our study we found that all the herbs showed reasonable % of Clotlysis.

These herbal extracts possess Thrombolytic properties that could lyse blood Clot s in vitro; however, in vivo Clot dissolving properties and active component(s) of these extracts for Clotlysis are yet to be discovered

Anti-Covid -19 potential of Azadirachta indica (Neem) leaf extract

Covid -19 is caused by infection with the “severe acute respiratory syndrome coronavirus-2″ (i.e., SARS-CoV-2). This is an enveloped virus having a positive sense, single-stranded RNA genome; like the two earlier viruses SARS-CoV and the Middle East respiratory syndrome (MERS) virus. Covid -19 is unique in that, in the severe case, it has the propensity to affect multiple organs, leading to multiple organ distress syndrome (MODS), and causing high morbidity and mortality in the extreme case. In addition, comorbidities like age, Cardiovascular Disease, diabetes and its complications, obesity, are risk factors for severe Covid -19.

It turns out that a most plausible, simple, single explanation for this propensity for MODS is the pivotal involvement of the vascular endothelium (VE). This is a consequence of the fact that the VE seamlessly connects all the entire vascular bed in the body, thus linking all the target organs (Heart, lungs, kidney, liver, brain) and systems.

Infection with SARS-CoV-2 leads to hyper-inflammation yielding uncontrolled production of a mixture of cytokines, chemokines, reactive oxygen species, nitric oxide, oxidative stress, acute phase proteins (e.g., C-reactive protein), and other pro-inflammatory substances. In the extreme case, a cytokine storm is created. Displacement of the virus bound to the VE, and/or inhibition of binding of the virus, would constitute an effective strategy for preventing Covid -19. In this regard, the acetone-water extract of the leaf of the Neem (Azadirachta indica) plant has been known to prevent the adherence of malaria parasitized red blood cells (pRBCs) to VE; prevent cytoadherence of cancer cells in metastasis; and prevent HIV from invading target T lymphocytes.

We therefore hypothesize that this Neem leaf acetone-water extract will prevent the binding of SARS-CoV-2 to the VE, and therefore be an effective therapeutic formulation against Covid -19. It is therefore advocated herein that this extract be investigated through rigorous clinical trials for this purpose. It has the advantages of being (i) readily available, and renewable in favor of the populations positioned to benefit from it; (ii) simple to prepare; and (iii) devoid of any detectable toxicity.

Antimicrobial and Thrombolytic Activities of Decoction of Azadirachta indica

The decoction of Azadirachta indica leaves were evaluated for their antimicrobial and Thrombolytic activities respectively. The antimicrobial activity was done by disk diffusion method where kanamycin (30μg/disc) was used as positive control (standard). The experiment showed highest activity against gram negative bacteria especially against E.coli and Vibrio parahaemolyticus and fungi particularly against Aspergillus niger. However, it was moderately active against gram positive bacteria.

For Thrombolytic activity, we used streptokinase as standard and normal saline and distilled water as negative control. In the test it has been observed that the decoction was moderately active for lysis of the blood Clot (27.53 ± 0.56)%. Our current studies for the first time, justified the use of decoctions of the leaves of Azadirachta indica for treating bacterial and fungal infection as well as use as thrombylitic agent in the indigenous system of medicine

Breviscapine (Erigeron Breviscapus Extract)

[Inhibitory effects of new-breviscapine on Thrombosis in vivo].

New-Breviscapine is an anti-platelet compound with some kinds of salts extracted from a Chinese herb Erigeron breviscapus. In aortic thrombus formation in rabbits, the platelet 5-HT release reaction and platelet destruction could be reduced by the compound, meanwhile aortic Thrombosis was inhibited with a clear correlation between drug dosage and its efficacy.

It was shown that new-breviscapine could inhibit the production of TXB2 and 6-keto-PGF1 alpha by platelets and endothelial cells respectively. The data suggest that new-breviscapine has a significant inhibitory effect on thrombus formation in vivo through suppression of platelet functions.

Protective effect of breviscapine in acute Pulmonary Embolism rats via regulation of MCP‐1 and IL‐13

Bre was shown to down‐regulate IL‐13 expression in the blood and further down‐regulate MCP‐1 expression, which caused alveolar macrophages (AMs) with lower cell activation efficiency.

The end result is to reduce inflammation of lung tissues, which facilitates the autologous Thrombolytic reaction and reduction in vasoconstriction in rats with PEs. Bre injections have been used >40 years, and may be recommended for the treatment of PE.

Additional clinical trials and data are warranted.

[Hypoxemia induced the changing structure of the lung tissue in SD rat though changing blood Clot ting and the effects of breviscapine’s intervention].

Objective: To investigate the expressions of plasminogen activator inhibitor-1 (PAI-1), activated protein C (APC) and the histology structures of the rat lung tissues in the different hypoxia time; and to investigate the effects of breviscapine to the above changes.

Methods: Eighty SD rats were randomly divided into A (control), B (hypoxia), C (hypoxia + low-dose breviscapine) and D (hypoxia + high-dose breviscopine) groups with 20 rats in each group. Each hypoxia group placed daily pressure (101 kpa, 10% O2) environment for 8 h, low-dose and high-dose breviscapine groups were given of 10 mg/kg, 40 mg/kg breviscapine by intraperitoneal injection. On the 3rd, 7th, 14th and 21st d, 5 rats were randomly taken from each group and were killed for examination. The hematoxylin and eosin stain (HE stain) was performed for observation on pathological changes in the rat lung tissues. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis was performed for detection of the mRNA levels of transforming growth factor (TGF-beta1) and Plasminogen activator inhibitor-1 (PAI-1). Western blot analysis was applied for detection of the expression of PAI-1. Besides, APC in the bronchoalveolar lavage fluid (BALF) was determined by ELISA.

Results: (1) The HE stain demonstrated that compared with A group, the degree of thickening of alveolar septal the mRNA expressions of TGF-beta1 and PA-1, and the protein expressions of PAI-1 in B group were increased (P < 0.05), and the expression of APC in the BALF was decreased (P < 0.05). And with prolonged hypoxia, the more significant of these changes were observed. Positive correlation was found between the mRNA levels of PAI-1 and TGF-beta1 (r = 0.936, P < 0.05). (2) Compared with B group, the increased thicknesses of alveolar septal in C and D groups were lightened, the mRNA expressions of TGF-beta1 and PAI-1, and the protein expression of PAI-1 were decreased (P < 0.05), and the expressions of APC in the BALF was increased (P < 0.05). With increasing dose, the expression levels of each factor gradually reduced or increased.

Conclusion: Hypoxia may cause coagulant function abnormality to increase Clotting activity and reduce fibrinolytic activity and the Anticoagulant activity, inducing alveolar septal thickening, and the mechanism of above changes may related to the TGF-beta1 signaling pathways. Breviscapine could improve hypoxia-induced hypercoagulable state that alleviate alveolar septal thickening.

therapeutic effects of Breviscapine in Cardiovascular Diseases : A Review

Breviscapine is a crude extract of several flavonoids of Erigeron breviscapus (Vant.) Hand.-Mazz., containing more than 85% of scutellarin, which has been traditionally used in China as an activating blood circulation medicine to improve cerebral blood supply.

Accumulating evidence from various in vivo and in vitro studies has shown that breviscapine exerts a broad range of Cardiovascular pharmacological effects, including vasodilation, protection against ischaemia/reperfusion (I/R), anti-inflammation, antiCoagulation, AntiThrombosis, endothelial protection, Myocardial protection, reduction of smooth muscle cell migration and proliferation, anticardiac remodeling, antiarrhythmia, blood lipid reduction, and improvement of erectile dysfunction.

In addition, several clinical studies have reported that breviscapine could be used in conjunction with Western medicine for Cardiovascular Diseases (CVDs) including Coronary Heart Disease, Myocardial Infarction, hypertension, Atrial Fibrillation, hyperlipidaemia, viral Myocarditis, chronic Heart failure, and Pulmonary Heart Disease.

However, the protective effects of breviscapine on CVDs based on experimental studies along with its underlying mechanisms have not been reviewed systematically. This paper reviewed the underlying pharmacological mechanisms in the cardioprotective effects of breviscapine and elucidated its clinical applications.

Bupleurum chinense DC. extract

Danhong Huayu Koufuye prevents Venous Thrombosis through antiinflammation via Sirtuin 1/NF-κB signaling pathway

Ethnopharmacological relevance: Danhong Huayu Koufuye (DHK), a compound traditional Chinese medicine, is composed of Salvia miltiorrhiza radix (Salvia miltiorrhiza Bge.), Angelicae Sinensis radix (Angelicae Sinensis (Oliv.) Diels.), Chuanxiong rhizoma (Ligusticum chuanxiong Hort.), Persicae semen (Prunus persica (L.) Batsch), Carthami flos (Carthamus tinctorius L.), Bupleuri radix (Bupleurum chinense DC.) and Aurantii fructus (Citrus aurantium L.). DHK prevents Deep vein Thrombosis (DVT) through antiinflammation . However, the antiinflammatory mechanism of DHK is still unknown.

Objective: The aim of this study was to evaluate whether DHK prevented Venous Thrombosis through antiinflammation via Sirtuin 1 (SIRT1)/NF-κB signaling pathway.

Methods: Inferior vena cava (IVC) stenosis-induced DVT rat model was established. Rats were administered with DHK (1.6, 3.2 or 6.4 mL/kg/d, p.o.), heparin (200 U/kg/d, i.v.), clopidogrel (25 mg/kg/d, p.o.), resveratrol (50 mg/kg/d, p.o.) or vehicle (p.o.) once daily for two days. blood Coagulation , blood fibrinolysis, blood viscosity, blood cell counts and platelet activity were evaluated. Serum levels of inflammatory cytokines were analyzed by enzyme-linked immunosorbent assay. Pathological changes were observed by hematoxylin-eosin (HE) staining. Protein expressions in thrombosed IVCs were evaluated by Western blot and/or immunofluorescence analyses. SIRT1 mRNA expression was analyzed by real-time quantitative polymerase chain reaction. Besides, SIRT1-specific inhibitor EX527 was pretreated to confirm the role of SIRT1/NF-κB signaling pathway in the antithrombotic effect of DHK.

Results: DHK remarkably prevented DVT. DHK had no effects on blood Coagulation , blood fibrinolysis, blood viscosity, blood cell counts or platelet activity. But DHK significantly up-regulated protein and mRNA expressions of SIRT1, and reduced leukocytes infiltration into thrombus and vein wall, serum levels of inflammatory cytokines, and protein expressions of acetylated p65 (Ace-p65), phosphorylated p65 (p-p65) and tissue factor (TF). Moreover, the antithrombotic effect of DHK was significantly abolished by EX527.

Conclusion: DHK may prevent DVT by inhibiting inflammation via SIRT1/NF-κB signaling pathway.

Potential Targets for Treatment of Coronavirus Disease 2019 (Covid-19 ): A Review of Qing-Fei-Pai-Du-Tang and Its Major Herbs

Covid -19 has been declared a pandemic by WHO on March 11, 2020. No specific treatment and vaccine with documented safety and efficacy for the Disease have been established. Hence it is of utmost importance to identify more therapeutics such as Chinese medicine formulae to meet the urgent need. Qing Fei Pai Du Tang (QFPDT), a Chinese medicine formula consisting of 21 herbs from five classical formulae has been reported to be efficacious on Covid -19 in 10 provinces in mainland China. QFPDT could prevent the progression from mild cases and shorten the average duration of symptoms and hospital stay. It has been recommended in the 6th and 7th versions of Clinical Practice Guideline on Covid -19 in China.

The basic scientific studies, supported by network pharmacology, on the possible therapeutic targets of QFPDT and its constituent herbs including Ephedra sinica, Bupleurum chinense, Pogostemon cablin, Cinnamomum cassia, Scutellaria baicalensis were reviewed. The anti-oxidation, immuno-modulation and antiviral mechanisms through different pathways were collated.

Two clusters of actions identified were cytokine storm prevention and angiotensin converting enzyme 2 (ACE2 ) receptor binding regulation. The multi-target mechanisms of QFPDT for treating viral infection in general and Covid -19 in particular were validated. While large scale clinical studies on QFPDT are being conducted in China, one should use real world data for exploration of integrative treatment with inclusion of pharmacokinetic, pharmacodynamic and herb-drug interaction studies.

platelet anti-aggregating activities of bupleurumin from the aerial parts ofBupleurum falcatum

An aryltetralin lactone, (-)-(1S,2R,3R)-1-(3’,4’-methylenedioxyphenyl)-3-(hydroxymethyl)-6,7- dimethoxy-1, 2,3,4-tetrahydronaphthalene-2-carboxylic acid lactone, was isolated from the methanol extract of the aerial parts ofBupleurum falcatum and named bupleurumin. The structure of the compound was determined on the basis of chemical and spectroscopic methods, including two-dimensional NMR spectrometry (e.g.,1H- and13C-NMR, homo- and heteronuclear COSY, HMBC).

Bupleurumin showed an 8-fold potent inhibitory effect (IC50: 47.5 μM) compared to that of acetylsalicylic acid (ASA, IC50: 420 μM) on collagen induced platelet aggregation and comparable effects as ASA on arachidonic acid-induced platelet aggregation.

Carthamus tinctorius L.FLOWER

Anti-thrombotic effect of Carthamus tinctorius Linn extracts in rats

Purpose: To explore the effects of Carthamus tinctorius L. (CTL) extracts on Thrombosis in rats.

Methods: CTL extract was obtained in hot water (60 ^oC), dried in a hot air oven and then freeze-dried.The rats were divided into 6 groups: normal group, control group, reference group (aspirin 5 mg/kg) as well as groups that received 20, 40 and 80 mg/kg doses of CTL, respectively. For each group, treatment was given orally once daily for 14 days. Common carotid Artery FeCl₃-induced thrombus and inferior vena cava Thrombosis occlusion time, as well as plasma concentrations of thromboxane B2 (TXB2) and 6-keto-prostaglandine F1α(6-keto-PGF1α) were measured in rats.

Results: Compared with the control group, all doses of CTL extracts significantly and dose-dependently prolonged Thrombosis occlusion time, reduced the weight of thrombus and increased inhibition rate (p < 0.01). Plasma TXB2 concentration of all CTL extracts groups decreased dose-dependently (p < 0.05) while that of 6-keto-PGF1α was increased (p < 0.05). There was association between 6-keto- PGF1α/TXB2 and Arterial or Venous thrombus weight for all treatments, and also with occlusion time for CTL treatment but not for aspirin.

Conclusion: CTL has a significant effect on Thrombosis in rats. However, further studies are required to determine its clinical potentials.

Assessment of Clot lytic effect of Carthamus tinctorius (Golrang).

Objectives: Carthamus tinctorius, also known as Kafeshe, Golrang and Kajirein Persian, is an herb from Compositea family and its seed is traditionally called Ghortom. Flowers of Carthamus tinctorius are usually used as a food flavoring agent.  Flower and oil of Carthamus tinctorius are considered as antithrombotic and anticoagulative agents in Iranian traditional medicine. In this study, the effects of different concentrations of Carthamus tinctorius extract were assessed on Clotlysis. Several in-vitro models have been developed to study Clot lytic activity of thrombolysis, but all of them have certain limitations. One of these limitations is hemolysis.

Materials and Methods: Whole blood from healthy individuals was allowed to form Clots in re-weighed sterile micro tubes; serum was removed and Clot was weighed. After lysis by seven different concentrationsof flow er extract (1.25 to 20%) fluid was removed and the remnants of the Clot were re-weighed with tubes. Percentage of Clotlysis was calculated on the basis of the weight difference of micro tubes obtained before and after Clotlysis . Also, upper fluid was used for hemolysis detecting by two methods: 1-potassium measurement 2-microscopy.

Results: The significant percentage of Clotlysis was observed when higher dosesof Carthamus tinctorius extracts were used (about 13.5%) in contrast with normal slain as a negative control (1-2%). Also no hemolysis was observed in all concentrations.

Conclusion: Carthamus tinctorius extract has Clot lytic activity in-vitro, furtheranimal studies are needed for confirming this idea.

Effect of the carthamins yellow from Carthamus tinctorius L. on hemorheological disorders of blood stasis in rats

Hemorheological disorders may play an important role in the pathogenesis and development of many Diseases . blood stasis, i.e. the decrease of blood flow velocity, indicates hemorheological abnormalities. The carthamins yellow (CY), isolated from Carthamus tinctorius L., has been extensively used as a natural food colorant. We investigated the effects of CY on a blood stasis model, which was obtained by placing rats in ice-cold water during the time interval between two injections of epinephrine. The results demonstrated that the CY significantly decreased the whole blood viscosity, plasma viscosity, and erythrocyte aggregation index, which were increased in the blood stasis model.

Hematocrit and platelet aggregation were reduced, while prothrombin time was delayed with increasing doses of CY. Therefore, CY administration might provide the additional benefit of increasing blood fluidity by lowering blood viscosity, which can be of great value in the prevention of hemorheological disorder-associated Diseases in at risk patients. Meanwhile, the mild activities of antiplatelet aggregation and antiCoagulation induced by CY should be considered, if these relatively untoward symptoms occurred when the hemorrhagic patients ate food colored by CY. However the small amounts used in food are highly unlikely to cause adverse effects.

Effect of Carthamus tinctorius L. (Safflower) on National Institute of Health Stroke Scale Scores of Ischemic Diseases Stroke Patients: A Pilot Clinical Trial

Experiencing complications within the first two weeks after Stroke leads to a high risk of mortality and length of hospitalization. The present pilot study was intended to investigate the hypothesis that adult patients treated with safflower or not would present with fewer neurological complications following 15 days. In a randomized controlled trial, subjects diagnosed with Ischemic Diseases cerebrovascular accident (CVA) based on focal neurological findings on brain imaging who met the inclusion criteria of our study were recruited from Ghaem Hospital, Mashhad, Iran, between 2016 and 2017.

Thirty-six patients were included in the survey and randomly allocated into treatment (A) and control (B) groups. An oral syrup of safflower extract and nasal drop of safflower oil were additionally prescribed for group A. Group B only received a standard anti-Ischemic Diseases regimen. The primary outcome measure was the National Institutes of Health Stroke Scale score (NIHSS) over 15 days.

Safflower treatment led to a notably higher mean difference in the NIHSS score between the baseline score and 15-day post-treatment score in group A in comparison to group B (p < 0.001). However, adjustment for covariates (age, gender, and baseline measures) showed no significant reduction in neurological status between them (p = 0.340). There was a statistically significant difference in neurological symptom scores between the groups (p = 0.044). Based on this pilot study, adjuvant treatment with safflow er in addition to the standard anti-Ischemic Diseases regimen can be more effective than individual conventional drugs for treating Ischemic Diseases CVA among adults.

Chaenomeles sinensis (Thouin) Koehne extract

Physiological Functionalities of Hot Water Extract and Some Medicinal Materials, and Their Mixtures

The aim of this study was to investigate the physiological functionalities of Codonopsis lanceolata, Glycyrrhiza uralensis, Chaenomeles sinensis, Crataegus pinnafida, and their mixtures. We also determined their antioxidative, fibrinolytic , and $\alpha$ inhibitory activities. The antioxidative activities of Codonopsis lanceolata, Glycyrrhiza uralensis, Chaenomeles sinensis, and Crataegus pinnafida were 79%, 88.3%, 89.9%, and 89.3% respectively. Their fibrinolytic activities were 0.80plasmin unit/mi, 0.57 plasmin unit/mi, 0.52 plasmin unit/mi, and 0.53 plasmin unit/mi respectively. The $\alpha$ inhibitory activity of Codonopsis lanceolata was 25%. The 10-fold diluents of Glycyrrhiza uralensis, Chaenomeles sinensis, and Crataegus pinnafida showed $\alpha$ inhibitory activities of 93.6%, 65.3%, and 61.3% respectively.

In antioxidative activity tests of the medicinal plants mixtures at various ratios, the mixtures of Glycyrrhiza uralensis, Chaenomeles sinensis, and Crataegus pinnafida with Codonopsis lanceolata showed antioxidative activities of approximately 90%. In fibrinolytic activity tests mixtures(1:1) of Codonopsis lanceolata with Chaenomeles sinensis and Crataegus pinnafida exhibited increases of 23% and 24% in activity respectively.

In $\alpha$ inhibitory activity tests, a mixture (4:1) of Codonopsis lanceolata and 10-fold diluted Glycyrrhiza uralensis showed an inhibitory activity of 98%, a mixture (3:1) of Codonopsis lanceolata and 10-fold diluted Chaenomeles sinensis showed an inhibitory activity of 69.6%, and a mixture (1:1) of Codonopsis lanceolata and 10-fold diluted Crataegus pinnafida showed an inhibitory activity of 50.2%. In conclusion, the mixtures of Glycyrrhiza uralensis, Chaenomeles sinensis, and Crataegus pinnafida with Codonopsis lanceolata will be used as a material for the development of biofunctional foods.

Cinnamomum cassia Presl. extract

The Extracts of Angelica sinensis and Cinnamomum cassia from Oriental Medicinal Foods Regulate Inflammatory and Autophagic Pathways against Neural Injury after Ischemic Diseases Stroke

The study indicates inflammation and autophagy are closely related to neural apoptosis in the pathology of Ischemic Diseases Stroke . In the study, we investigate the effects and mechanisms of the extracts of Angelica sinensis and Cinnamomum cassia (AC) from oriental medicinal foods on inflammatory and autophagic pathways in rat permanent middle cerebral Artery occlusion model.

Three doses of AC extract were, respectively, administered for 7 days. It suggests that AC extract treatment ameliorated scores of motor and sensory functions and ratio of glucose utilization in thalamic lesions in a dose-dependent manner. Expression of Iba1 was decreased and CD206 was increased by immunofluorescence staining, western blotting results showed expressions of TLR4, phosphorylated-IKKβ and IκBα, nuclear P65, NLRP3, ASC, and Caspase-1 were downregulated, and Beclin 1 and LC3 II were upregulated. Low concentrations of TNF-α, IL-1β, and IL-6 were presented by ELISA assay. Additionally, caspase 8 and cleaved caspase-3 expressions and the number of TUNEL positive cells in ipsilateral hemisphere were decreased, while the ratio of Bcl-2/Bax was increased.

Simultaneously, in LPS-induced BV2 cells, it showed nuclear P65 translocation and secretion of proinflammatory cytokines were suppressed by AC extract-contained cerebrospinal fluid, and its intervened effects were similar to TLR4 siRNA treatment. Our study demonstrates that AC extract treatment attenuates inflammatory response and elevates autophagy against neural apoptosis, which contributes to the improvement of neurological function postStroke.

Therefore, AC extract may be a novel neuroprotective agent by regulation of inflammatory and autophagic pathways for Ischemic Diseases Stroke treatment.

Anti-coagulant activity of plants: mini review

Medicinal plants have been used for treatment of human ailments since ancient times. Objective of this study is to document the effect of herbal drugs on Anticoagulant therapy. The material for this review was taken mostly from PubMed and the Cochrane database of systematic reviews. Some other relevant references were collected from personal database of papers on anti-coagulant properties of plants.

Literature review shows that many plants such as Thymus vulgaris, Cyamopsis tetragonoloba taub, Pulmonaria officinalis and 💥Cinnamomum cassia etc have anti-coagulant activity. This review shows that medicinal plants should be prescribed with care to patients on Anticoagulant therapy.

Citrus aurantium L. extract

In-vitro Assessment of Cholinesterase Inhibitory and Thrombolytic Activity of Six Available Citrus Fruits in Bangladesh: Relevant for Treating Neurodegenerative Disorder

Aim: Citrus fruits are well known for its medicinal and food value. Aim of this study is to investigate acetylcholinesterase ((AChE)) inhibitory activity, butyrylcholinesterase (BuChE) inhibitory activity, total phenolics, flavonoids, flavonols content and Thrombolytic activities of crude methanol extracts of 6 citrus fruits (Citrus limon, Citrus aurantifollia, Citrus bergamia, Citrus maxima, Citrus sinensis and Citrus macroptera).

Methods: The fruits were extracted by using methanol as solvent. Ellman’s colourimetric method was applied to determine both cholinesterase inhibitory activities, while folin-ciocalteau reagent (FCR) and aluminium chloride were used to quantify total phenolics, flavonoids, flavonol content of those fruits. blood Clotlysis method was applied for determining the Thrombolytic activity of those fruits.

Results: All citrus fruits contain a good amount of phenolics, flavonoids and flavonols. C. maxima found more prominent in containing phenolics and flavonols compare to other citrus fruits, with 414.06 ± 2.87 mg Gallic Acid Equivalent/gm and 12.94 ± 1.31 mg Catechin Equivalent/gm dried extract respectively. Citrus sinensis showed the highest content in flavonoids with 21.16± 1.37 mg Catechin 20 Equivalent /gm dried extract. Citrus fruits are also a quality source of cholinesterase inhibitors. All the examined citrus fruits were found capable of inhibiting both acetylcholinesterases (AChE) as well as butyrylcholinesterase (BuChE). C. bergamia was most effective in inhibiting AChE with IC50 of 27.18 μg/ml where C. macroptera was best in inhibiting BuChE (IC50 32.5 μg/ml). But none of the citrus fruits was found fit for Thrombolytic activity.

Conclusion: Citrus fruits are found the sound in inhibiting AChE and BuChE as well as containing Phenolics, flavonoids and flavonols. But they lack in their Thrombolytic activity.

An overview of Citrus aurantium used in treatment of various Diseases

Citrus aurantium (bitter orange) is a plant belonging to the family Rutaceae, The most important biologically active constituents of the C. aurantium fruits are phenethylamine alkaloids octopamine, synephrine, tyramine, N-methyltyramine and hordenine. It is rich in vitamin C, flavonoids and volatile oil.

Synephrine is a primary synthesis compound with pharmacological activities such as vasoconstriction , elevation of blood pressure and relaxation of bronchial muscle. whose fruit extracts have been used for the treatment of various Diseases such as gastrointestinal disorders, insomnia, head aches, Cardiovascular Diseases, cancer, antiseptic, anti-oxidant, antispasmodic, aromatic, astringent, carminative, digestive, sedative, stimulant, stomachic and tonic and by research novel use
is found in obesity and related risks even life threatening are continuously increasing through out world in all age groups.

Many marketed formulations claim to possess antiobesity actions, but still many herbs which have claims to this need to be investigated and their claims to be authenticated. In recent era there is a great thrust on screening of herbal extracts and formulations for antiobesity action. In this article efforts have been taken to discuss the photochemistry, constituents, ethnobotany, pharmacology safety and toxicity of citrus plant. The motto is to discuss C. aurantium here more research attention should be given on this that would increase its use in various chronic and acute Diseases.

Antiobesity and Cardiovascular toxic effects of Citrus aurantium extracts in the rat: a preliminary report

Danhong Huayu Koufuye prevents Venous Thrombosis through antiinflammation via Sirtuin 1/NF-κB signaling pathway

Abstract

Ethnopharmacological relevance: Danhong Huayu Koufuye (DHK), a compound traditional Chinese medicine, is composed of Salvia miltiorrhiza radix (Salvia miltiorrhiza Bge.), Angelicae Sinensis radix (Angelicae Sinensis (Oliv.) Diels.), Chuanxiong rhizoma (Ligusticum chuanxiong Hort.), Persicae semen (Prunus persica (L.) Batsch), Carthami flos (Carthamus tinctorius L.), Bupleuri radix (Bupleurum chinense DC.) and Aurantii fructus (💥Citrus aurantium L.). DHK prevents Deep vein Thrombosis (DVT) through antiinflammation . However, the antiinflammatory mechanism of DHK is still unknown.

Objective: The aim of this study was to evaluate whether DHK prevented Venous Thrombosis through antiinflammation via Sirtuin 1 (SIRT1)/NF-κB signaling pathway.

Methods: Inferior vena cava (IVC) stenosis-induced DVT rat model was established. Rats were administered with DHK (1.6, 3.2 or 6.4 mL/kg/d, p.o.), heparin (200 U/kg/d, i.v.), clopidogrel (25 mg/kg/d, p.o.), resveratrol (50 mg/kg/d, p.o.) or vehicle (p.o.) once daily for two days. blood Coagulation , blood fibrinolysis, blood viscosity, blood cell counts and platelet activity were evaluated. Serum levels of inflammatory cytokines were analyzed by enzyme-linked immunosorbent assay. Pathological changes were observed by hematoxylin-eosin (HE) staining. Protein expressions in thrombosed IVCs were evaluated by Western blot and/or immunofluorescence analyses. SIRT1 mRNA expression was analyzed by real-time quantitative polymerase chain reaction. Besides, SIRT1-specific inhibitor EX527 was pretreated to confirm the role of SIRT1/NF-κB signaling pathway in the antithrombotic effect of DHK.

Results: DHK remarkably prevented DVT. DHK had no effects on blood Coagulation , blood fibrinolysis, blood viscosity, blood cell counts or platelet activity. But DHK significantly up-regulated protein and mRNA expressions of SIRT1, and reduced leukocytes infiltration into thrombus and vein wall, serum levels of inflammatory cytokines, and protein expressions of acetylated p65 (Ace-p65), phosphorylated p65 (p-p65) and tissue factor (TF). Moreover, the antithrombotic effect of DHK was significantly abolished by EX527.

Conclusion: DHK may prevent DVT by inhibiting inflammation via SIRT1/NF-κB signaling pathway.

Citrus assamensis

Screening of Antibacterial, Thrombolytic , Membrane Stabilizing, anti-inflammatory and Antitumor Activity of Citrus assamensis Leaf Extracts

The methanol, ethanol and chlorofom leaf extracts of Satkara, Citrus assamensis (family: Rutaceae), were subjected to in vitro anti-bacterial, Thrombolytic , membrane stabilizing and in vivo anti-inflammatory and antitumor activity tests. The chloroform extract of C. assamensis showed the most important spectrum of activity against Bacillus subtilis, Bacillus cereus, Sarcina lutea among 6 gram positive and against 11 gram negative bacteria at the concentration of 1000 μg/disc, while the range of zones of inhibition were within 7-16 mm. Among the tested three extracts CHCl₃ extract showed potent Thrombolytic activity and hypotonic solution induced haemolytic activity where the percentages of inhibition were found to be 35% and 55% respectively. All the extracts established significant (p<0.05) anti-inflammatory effect by regulating biphasic inflammatory process induced by carrageenan. The leaf extract dose-dependently and significantly decreases the number of EAC cell count and inhibition of cell growth in comparison to the EAC control and standard. The results obtained in the present study indicate that, C. assamensis leaf can be a potential source of anti-bacterial, Thrombolytic , membrane stabilizing, anti-inflammatory and antitumor agent s.

Phytochemical Screenings and Anti-Thrombolytic Activity of Citrus Assamenses

In this present study, the leaf extracts of Citrus assamenses were subjected to evaluate preliminary phytochemical investigation and Thrombolytic activities The Thrombolytic activity were assessed by using human blood and the results were compared with standard streptokinase (SK). The present study showed that the carbon tetra chloride soluble fraction (CSF) exhibited highest Thrombolytic activity (39.82%). However, significant Thrombolytic activity was demonstrated by the ethanol soluble fraction (ESF) of C. assamenses (35.95%), where as the positive control showed 93.29% lysis of Clot . Preliminary phytochemical investigation suggested the presence of alkaloids, tannins, reducing sugars and flavonoids.

Clerodendrum viscosum (Verbanaceae)

Evaluation of Thrombolytic potential of three medicinal plants available in Bangladesh, as a potent source of Thrombolytic compounds

Objective: The present study is aimed to investigate in vitro Thrombolytic activity of three Bangladeshi medicinal plants Averrhoa bilimbi (Oxalidiaceae), Clerodendrum viscosum (Verbanaceae) and Drynaria quercifolia (Polypodiaceae).

Materials and methods: Each the plant was extracted with methanol at room temperature and the concentrated methanolic extracts (MEF) were fractionated by the modified Kupchan partitioning method to render pet-ether soluble fraction (PESF), carbon tetrachloride soluble fraction (CTSF), chloroform soluble fraction (CSF) and aqueous soluble fraction (AQSF). To observe their Thrombolytic potential, a prompt and swift method was involved where streptokinase and water were used as positive and negative control, respectively.

Result: Among the three plants, AQSF and PESF of D. quercifolia with CTSF of C. viscosum exhibited highest Thrombolytic activity by Clotlysis of 34.38%, 34.27% and 28.64%, respectively. Among other extracts A. bilimbi, C. viscosun and D.quercifolia showed significant percentage (%) of Clotlysis compared to standard streptokinase (41.05%) while the negative control water revealed 3.31 % lysis of Clot .

Conclusion: From our findings it is observed that all the plants revealed remarkable Thrombolytic activity. Therefore, steps should be taken to observe in vivo Clot dissolving potential and to isolate active component(s) of these extracts.

Naturally Growing Medicinal Plant Clerodendrum viscosum in Bangladesh – Thrombolytic , Membrane Stabilizing and Anti-microbial properties .

Roots of Clerodendrum viscosum are profoundly used throughout the world due to its ethno-medicinal properties and folkloric use since the very beginning of medicinal treatment. Due to the potential Disease preventive properties , we investigate the methanolic extracts of the roots of the plant for biological evaluation in vitro.The methanolic extract of Cl. viscosum roots was partitioned into ethyl acetate soluble fraction (ESF), petroleum ether soluble fraction (PSF), carbon tetrachloride soluble fraction (CTSF), chloroform soluble fraction (CSF) and aqueous soluble fraction (AQSF). The extracts were evaluated for their Thrombolytic , membrane stabilizing, antimicrobial and the results were compared with standard drugs; streptokinase for Thrombolytic , acetyl salicylic acid for anti-inflammatory , kanamycin for antimicrobial activities. In Thrombolytic investigation, among all partitionates, the ESF showed highest % of Clotlysis (54.47%) as compared to (69.13%) and (3.17%) exhibited by the standard streptokinase and water. In case of membrane stabilizing study, ESF also significantly inhibited the haemolysis of human erythrocyte membrane both induced by hypotonic solution (60.30 ± 0.54%) and by heat (44.21 ± 0.59%), respectively as compared to (69.01 ± 16%) and (71.90 ± 0.19%) demonstrated by acetyl salicylic acid. In antibacterial activity, it was exhibited by the extracts, was comparatively more prominent on the gram negative bacteria than the gram positive bacteria. Our study revealed that satisfactory amount of flavonoid and tannin presence showed a significant and positive correlation between bioactive compound contents with pharmacological activities of Cl. Viscosum roots.

Characterization of active Anticoagulant fraction and a fibrin(ogen)olytic serine protease from leaves of Clerodendrum colebrookianum, a traditional ethno-medicinal plant used to reduce hypertension

Ethnopharmacological relevance

Cardiovascular Diseases are the major cause of mortality and morbidity, causing over 17.9 million deaths a year worldwide. Currently used therapy is often having side effects and expensive, dietary interventions and alternative medicines are required. Clerodendrum colebrookianum has been used to treat cardiac hypertension but Anticoagulant potency was not evaluated.
Aim of the study

To characterize an active Anticoagulant fraction (AAFCC) and a 30 kDa fibrin(ogen)olytic serine protease (clerofibrase) isolated from aqueous leave extract of C. colebrookianum.
Materials and methods

AAFCC/clerofibrase was subjected to extensive biochemical and pharmacological characterization including LC-MS/MS, amino acid compositional and GC-MS analyses. Interaction between clerofibrase with fibrinogen was studied by spectrofluorometric analysis. In vitro Thrombolytic , antiplatelet and cytotoxicity assay were performed. In vivo toxicity, Anticoagulant , defibrinogen and antithrombotic activities were determined on Swiss albino mice.
Results

The in vitro Anticoagulant activity of AAFCC was found to be superior to heparin and clerofibrase and comparable to Nattokinase and warfarin. The proteomics and amino acid composition analyses suggest that clerofibrase is a previously uncharacterized novel plant protease capable of degrading the -αβ chains of fibrinogen/fibrin. AAFCC/clerofibrase exerts their Anticoagulant action via fibrinogenolytic activity and partially by antiplatelet activity albeit they have no effect on thrombin and FXa inhibition . The spectrofluorometric analysis revealed the binding of clerofibrase to fibrinogen but not to thrombin and FXa. The phytochemical constituents and bioactive components of AAFCC were characterized by biochemical, and GC-MS analyses. The AAFCC and clerofibrase inhibited collagen/ADP-induced mammalian platelet aggregation , showed in vitro Thrombolytic activity, and non-cytotoxic to mammalian cells. The AAFCC showed and dose-dependent in vivo plasma defibrinogenating and Anticoagulant activities and inhibited k-carrageen-induced thrombus formation in the tails of mice.
Conclusion

The potent in vivo Anticoagulant and antithrombotic effects of AAFCC suggests its pharmacological significance as herbal Anticoagulant drug for the prevention and/or treatment of hyperfibrinogenemia- and Thrombosis associated Cardiovascular disorders .

Antihemolytic activity of Clerodendrum viscosum Vent. is mediated by its antioxidant effect.

Aims: The objective of this study was to evaluate the protective efect of polar extract of Clerodendrum viscosum vent. against in vitro hemolysis of human erythrocyte s and its asociation with the antioxidant activity of C. viscosum. Study Design: Extraction of C. viscosum dried rot, in vitro antihemolytic activity asay, lipid peroxidation asay, phytochemical analysis, estimation of polyphenols and flavonoids. Place and Duration of Study: Department of Biochemistry and Molecular Biology, Jahangirnagar University, Savar, Dhaka, Bangladesh, betwen January 201 and May 201.

Methodology: C. viscosum polar extract-pretreated erythrocytes were hemolysed by hypotonia and oxidizing agent, H2O2. The liberated hemoglobin was determined as a measure of hemolysis. Total reducing power and 2,2-diphenyl-1-picrylhydrazyl (DPPH)- fre radical scavenging activity of extract were compared with those of vitamin C. Antilipid peroxidation activity of C. viscosum polar extract also was determined by exposing rat brain cortex tisue to Fenton’s reagent (H2O2+FeSO4)- induced oxidative stres. Then C. viscosum extract was subjected to estimation of total polyphenols and total flavonoids folowing qualitative phytochemical analysis.

Results: C. viscosum polar extract signifcantly inhibited in vitro hemolysis. Total reducing power and DPPH-radical scavenging activity were higher than those of vitamin C. In cortical tisue homogenate, C. viscosum polar extract signifcantly reduced (~38%) the levels of lipid peroxides (LPO). Phytochemical analysis revealed the presence of substantial amount of polyphenols, flavonoids and other antioxidant chemicals in the extract.

Conclusion: Present investigation demonstrates that antihemolytic activity of C. viscosum polar extract is mediated by its antioxidant effect.

Coicis Semen extract

Coicis semen protects against focal cerebral ischemia –reperfusion injury by inhibiting oxidative stress and promoting angiogenesis via the TGFβ/ALK1/Smad1/5 signaling pathway

cordycepin

anti-inflammatory effects of cordycepin: A review

Cordycepin is the major bioactive component extracted from Cordyceps militaris. In recent years, cordycepin has received increasing attention owing to its multiple pharmacological activities. This study reviews recent researches on the anti-inflammatory effects and the related activities of cordycepin. The results from our review indicate that cordycepin exerts protective effects against inflammatory injury for many Diseases including acute lung injury (ALI), asthma, rheumatoid arthritis, Parkinson’s Disease (PD), hepatitis, atherosclerosis, and atopic dermatitis. Cordycepin regulates the NF-κB, RIP2/Caspase-1, Akt/GSK-3β/p70S6K, TGF-β/Smads, and Nrf2/HO-1 signaling pathways among others. Several studies focusing on cordycepin derivatives were reviewed and found to down metabolic velocity of cordycepin and increase its bioavailability.

Moreover, cordycepin enhanced immunity, inhibited the proliferation of viral RNA, and suppressed cytokine storms, thereby suggesting its potential to treat Covid -19 and other viral infections. From the collected and reviewed information, this article provides the theoretical basis for the clinical applications of cordycepin and discusses the path for future studies focusing on expanding the medicinal use of cordycepin. Taken together, cordycepin and its analogs show great potential as the next new class of anti-inflammatory agents.

Cordycepin: a bioactive metabolite of C ordyceps militaris and polyadenylation inhibitor with therapeutic potential against Covid -19

Spike protein and main proteases of SARS-CoV-2 have been identified as potential therapeutic targets and their inhibition may lead to the reticence of viral entry and replication in the host body. Despite several efforts; till now no specific drugs are available to treat SARS-CoV-2 . Considering all these challenges, the main objective of the present study was to establish therapeutic potential of cordycepin against Covid -19 as a conventional therapeutic strategy. In the present study; molecular interaction study was performed to assess potential binding affinity of cordycepin with SARS-CoV-2 target proteins using computational approach.

Additionally, network pharmacology was used to understand cordycepin-protein interactions and their associated pathways in human body. Cordycepin is under clinical trial (NCT00709215) and possesses structural similarity with adenosine except that, it lacks a 3′ hydroxyl group in its ribose moiety and hence it served as a poly(A) polymerase inhibitor and terminate premature protein synthesis. Additionally, it is known that functional RNAs of SARS-CoV-2 genome are highly 3′-plyadenylated and leading to synthesis of all viral proteins and if cordycepin can destabilize SARS-CoV-2 RNAs by inhibiting polyadenylation process then it may step forward in terms of inhibition of viral replication and multiplication in the host.

Moreover, cordycepin showed strong binding affinity with SARS-CoV-2 spike protein (-145.3) and main proteases (-180.5) that further corroborate therapeutic potential against Covid -19. Since cordycepin has both pre-clinical and clinical information about antiviral activities, therefore; it is suggested to the world community to undertake repurposing cordycepin to test efficacy and safety for the treatment of Covid -19.

Potent Inhibitory Activities of the Adenosine Analogue Cordycepin on SARS-CoV-2 Replication

Nucleoside analogues are among the most successful bioactive classes of druglike compounds in pharmaceutical chemistry as they are well-known for their numerous effective bioactivities in humans, especially as antiviral and anticancer agent s. Coronavirus Disease 2019 (Covid -19 ) is still untreatable, with its causing virus, the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2 ), continuing to wreak havoc on the ground everywhere.

This complicated international situation urged all concerned scientists, including medicinal chemists and drug discoverers, to search for a potent anti-Covid -19 drug. Cordycepin (3′-deoxyadenosine) is a known natural adenosine analogue of fungal origin, which could also be synthetically produced. This bioactive phytochemical compound is characterized by several proven strong pharmacological actions that may effectively contribute to the comprehensive treatment of Covid -19 , with the antiviral activities being the leading ones. Some new studies predicted the possible inhibitory affinities of cordycepin against the principal SARS-CoV-2 protein targets (e.g., SARS-CoV-2 spike (S) protein, main protease (M^pro) enzyme, and RNA-dependent RNA polymerase (RdRp) enzyme) based on the computational approach. Interestingly, the current research showed, for the first time, that cordycepin is able to potently inhibit the multiplication of the new resistant strains of SARS-CoV-2 with a very minute in vitro anti-SARS-CoV-2 EC₅₀ of about 2 μM, edging over both remdesivir and its active metabolite GS-441524.

The ideal pharmacophoric features of the cordycepin molecule render it a typical inhibitor of SARS-CoV-2 replication, with its flexible structure open for most types of derivatization in the future. Briefly, the current findings further support and suggest the repurposing possibility of cordycepin against Covid -19 and greatly encourage us to confidently and rapidly begin its preclinical/clinical evaluations for the comprehensive treatment of Covid -19.

Cordycepin as a Promising Inhibitor of SARS-CoV-2 RNA Dependent
RNA Polymerase (RdRp)

Background: SARS-CoV-2 , which emerged in Wuhan, China, is a new global threat that has killed millions of people and continues to do so. This pandemic has not only threatened human life but has also triggered economic downturns across the world. Re- searchers have made significant strides in discovering molecular insights into SARS- CoV-2 pathogenesis and developing vaccines, but there is still no successful cure for SARS-CoV-2 infected patients.

Objective: The present study has proposed a drug-repositioning pipeline for the design and discovery of an effective fungal-derived bioactive metabolite as a drug candidate against SARS-CoV-2.

Methods: Fungal derivative “Cordycepin” was selected for this study to investigate the inhibitory properties against RNA-dependent RNA polymerase (RdRp) (PDB ID: 6M71) of SARS-CoV-2 . The pharmacological profile, intermolecular interactions, binding energy, and stability of the compound were determined utilizing cheminformatic approaches. Subsequently, molecular dynamic simulation was performed to better understand the binding mechanism of cordycepin to RdRp.

Results: The pharmacological data and retrieved molecular dynamics simulations trajectories suggest excellent drug-likeliness and greater structural stability of cordycepin, while the catalytic residues (Asp760, Asp761), as well as other active site residues (Trp617, Asp618, Tyr619, Trp800, Glu811) of RdRp, showed better stability during the overall simulation span.

Conclusion: Promising results of pharmacological investigation along with molecular simulations revealed that cordycepin exhibited strong inhibitory potential against SARS-CoV-2 polymerase enzyme (RdRp). Hence, cordycepin should be highly recommended to test in a laboratory to confirm its inhibitory potential against the SARS-CoV-2 polymerase enzyme (RdRp).

Possible pharmacological mechanisms of adenosine receptor and cordycepin from Cordyceps militaris in the prevention and treatment of Covid -19 pneumonia

Through the activation of adenosine receptors A1, A2A, A2B, and A3, adenosine plays an important role in the pathol. process of acute lung injury, such as anti-inflammatory, inhibiting cytokine storm, protecting organ damage, and repairing and remodeling tissues. cordycepin is also an activator of adenosine receptors.

It can enhance human immunity, inhibit RNA virus reproduction, anti-inflammatory, inhibit cytokine storm, protect liver, Heart, kidney, and lung fibrosis in clin. or animal models. Cordyceps and cordycepin, as a safe food supplement and(or) potential adenosine receptor agonist drug, can play a pos. role in the prevention and treatment of Covid -19 pneumonia.

Interactions Between Adenosine Receptors and Cordycepin from Cordyceps Militaris: Possible Pharmacological Mechanisms for Protection of the Brain and the Amelioration of Covid -19 Pneumonia

At present, the novel Covid -19 pneumonia is prevalent, affecting millions of people. Here, we summarized the pharmacological basis of adenosine, adenosine receptors, adenosine agonist cordycepin (3&#39;-deoxyadenosine), and Cordyceps product in the brain protection and amelioration of pneumonia to provide useful information to cope with the global pandemic of novel coronavirus (Covid -19 ). Adenosine, a mediator of innate immunity, is abundantly secreted by the injured lung tissues during inflammation.

Through the activation of adenosine receptors A1, A2A, A2B and A3, adenosine plays an important role in protecting against acute lung injury and brain injury. Cordycepin (3-deoxyadenosine) is an activator of adenosine receptors. It can enhance human immunity, promote anti-inflammatory processes, inhibit RNA virus reproduction, protect against brain, lung, liver, Heart, and kidney damage, and ameliorate lung-fibrosis in clinical and animal models. Cordyceps and cordycepin products could be used as a potential medicinal adenosine receptor agonist that can play a beneficial role in the amelioration of Covid -19 pneumonia and protection of brain.

Cordyceps sinensis (Berk.) Sacc extract

A novel extracellular protease with fibrinolytic activity from the culture supernatant of Cordyceps sinensis: purification and characterization

A novel serine protease with fibrinolytic activity named CSP was purified from the culture supernatant of the fungus Cordyceps sinensis, a kind of Chinese herbal medicine. Analysis of the purified enzyme by SDS-PAGE indicated that CSP was a single polypeptide chain with an apparent molecular weight of 31 kDa, and N-terminal sequencing revealed that the first ten amino acid residues of the enzyme were Ala-Leu-Ala-Thr-Gln-His-Gly-Ala-Pro-Trp-. When casein was used as a substrate, the proteolytic activity of CSP reached its maximum at pH 7.0 and 40 °C. The effect of chemical agent s on the enzyme activity indicated that CSP is a serine protease with a free cysteine residue near the active site.

It hydrolysed fibrinogen, fibrin and casein with a high efficiency, while hydrolysing bovine serum albumin (BSA) and human serum albumin (HSA) to a lesser extent. CSP was found to be a plasmin-like protease, but not a plasminogen activator, and it preferentially cleaved the Aα chain of fibrinogen and the α-chain of fibrin. Therefore, the extracellular protein CSP may represent a potential new therapeutic agent for the treatment of Thrombosis.

Biochemical characterization of a novel fibrinolytic enzyme from Cordyceps militaris

A fibrinolytic enzyme was produced by the medicinal mushroom, Cordyceps militaris using submerged fermentation. The enzyme was purified from culture supernatant by hydrophobic interaction, ion exchange and gel filtration chromatographies. It was purified by 36 fold, with a specific activity of 1,467.4 U/mg protein and the final yield was 5.8%. The molecular weight of the enzyme as determined by SDS-PAGE and gel filtration was 28 kDa and 24.5 kDa, respectively, and its isoelectric point (pI) was 9.0 ± 0.2. It was found to be a glycoprotein with carbohydrate content of 1.67% (w/v).

The enzyme was optimally active at 37 °C and pH 7.2. The enzyme activity was strongly inhibited by soybean trypsin inhibitor (SBTI) and aprotinin which indicated it to be a serine protease, while other inhibitors like N-α-tosyl-l-phenylalanine chloromethyl ketone (TPCK), phenyl methane sulfonyl fluoride (PMSF), pepstatin and metal chelator EDTA did not inhibit its activity. Amino acid sequences of the purified enzyme were determined partially by Q-TOF2 and they were IEDFPYQVDLR; ANCGGTVISEK; YVLTAGHCAEGYTGLNIR; TNYASVTPITADMICAGFPEGK; KDSCSGDSGGPLVTGGK; VVGIVSFGTGCAR; ANKPGVYSSVASAEIR.

Sequences of the seven peptides completely matched with those of a trypsin-like serine protease from Cordyceps militaris CM01 (accession no. EGX95217.1). The purified enzyme degraded α chains of fibrinogen first and then β and γ chains and also activated plasminogen into plasmin. It can act as an Anticoagulant and prevent Clot formation by degrading fibrinogen. Based on these studies, the purified enzyme has great potential to be developed as a natural agent for prevention and treatment of Thrombolytic Diseases.

A novel fibrinolytic enzyme from Cordyceps militaris, a Chinese traditional medicinal mushroom

A novel fibrinolytic enzyme from Cordyceps militaris was purified and partially characterized for the first time, which was designated C. militaris fibrinolytic enzyme (CMase). This extracellular enzyme from C. militaris was isolated by ammonium sulphate fraction, and purified to electrophoretic homogeneity using gel filtration chromatography. The apparent molecular mass of the purified enzyme was estimated to be 27.3 kDa by SDS-PAGE.

The optimum pH and temperature for the enzyme activity were pH 6.0 and 25 °C, respectively. In the presence of metal ions such as Mg2+ and Fe2+ ions the activity of the enzyme increased, whereas EDTA and Cu2+ ion inhibited the enzyme activity. Interestingly the N-terminal amino acid sequences of the enzyme is extremely similar to those of the trypsin proteinases from insects, and has no significant homology with those of the fibrinolytic enzyme from other medicinal mushroom. In conclusion, C. militaris produces a strong fibrinolytic enzyme CMase and may be considered as a new source for Thrombolytic agents.

A fibrinolytic Enzyme from the Medicinal Mushroom Cordyceps militaris

In this study we purified a fibrinolytic enzyme from Cordyceps militaris using a combination of ion-exchange chromatography on a DEAE Sephadex A-50 column, gel filtration chromatography on a Sephadex G-75 column, and FPLC on a HiLoad 16/60 Superdex 75 column. This purification protocol resulted in a 191.8-fold purification of the enzyme and a final yield of 12.9 %. The molecular mass of the purified enzyme was estimated to be 52 kDa by SDS-PAGE, fibrin-zymography, and gel filtration chromatography.

The first 19 amino acid residues of the N-terminal sequence were ALTTQSNV THGLATISLRQ, which is similar to the subtilisin-like serine protease PR1J from Metarhizium anisopliae var. anisopliase. This enzyme is a neutral protease with an optimal reaction pH and temperature of 7.4 and ${37}^{\circ }C$, respectively. Results for the fibrinolysis pattern showed that the enzyme rapidly hydrolyzed the fibrin $\alpha$-chain followed by the $\gamma$–$\gamma$ chains. It also hydrolyzed the $\beta$-chain, but more slowly. The A$\alpha$, B$\beta$, and $\gamma$ chains of fibrinogen were also cleaved very rapidly. We found that enzyme activity was inhibited by $C{u}^{2+}$ and $C{o}^{2+}$, but enhanced by the additions of $C{a}^{2+}$ and $M{g}^{2+}$ ions.

Furthermore, fibrinolytic enzyme activity was potently inhibited by PMSF and APMSF. This enzyme exhibited a high specificity for the chymotrypsin substrate S-2586 indicating it’s a chymotrypsin-like serine protease. The data we present suggest that the fibrinolytic enzyme derived from the edible and medicinal mushroom Cordyceps militaris has fibrin binding activity, which allows for the local activation of the fibrin degradation pathway.

Integrative medicine considerations for convalescence from mild-to-moderate Covid -19 Disease

The majority of individuals infected with SARS-CoV-2 have mild-to-moderate Covid -19 Disease . Convalescence from mild-to-moderate (MtoM) Covid -19 Disease may be supported by integrative medicine strategies. Integrative Medicine (IM) is defined as healing-oriented medicine that takes account of the whole person, including all aspects of lifestyle. Integrative medicine strategies that may support recovery from MtoM Covid -19 are proposed given their clinically studied effects in related conditions. Adoption of an anti-inflammatory diet, supplementation with vitamin D, glutathione, melatonin, Cordyceps, Astragalus and garlic have potential utility.

Osteopathic manipulation, Qigong, breathing exercises and aerobic exercise may support Pulmonary recovery. Stress reduction, environmental optimization, creative expression and aromatherapy can provide healing support and minimize enduring trauma. These modalities would benefit from clinical trials in people recovering from Covid -19 infection.

Curcumae Radix extract

Etiology and Chinese Medicine Screening of Severe Covid -19 Based on Multiomics and Serum Pharmacology

Background: At present, scholars believe that severe Covid -19 is related to a variety of basic Diseases , and we also observe this point using multi-omics method. The latest sequencing data of severe Covid -19 patients were combined to analyze the pathological mechanism, and pharmacological experimental research on local drugs was conducted, and a compound ingredient was found to have potential medicinal value.

Results: Here, we observed, for patients with severe Covid -19 Disease , the differential miRNA expression is mainly low but having higher expression of mRNA. These differential mRNA expressions are associated with the activation of inflammatory pathways and ultimately with hypoxia and Coagulation . Using database analysis, we found that Yi Xin Tong Mai Granule(YXTMG) might regulate Covid -19 through Toll-like receptor signaling pathway by acting on different immune targets. We found a new molecular mechanism for Covid -19 to turn the crisis around, the down-regulated miR-181a-5p mediates the up- regulation of PLAU and SERPINE1 molecules to cause Cardiovascular adverse events, and YXTMG may prevent it. At the same time, molecular docking indicated that the its various components have anti-inflammatory activity. In vitro studies, we confirmed that YXTMG had antioxidant and anti-inflammatory activities.

Conclusions: The study has supplemented the potential mechanism for the conversion of mild to critical Covid -19 Disease and screened the Chinese medicines for improving these factors, providing methodological reference for Disease pathology and drug development.

Curcumae Radix: a review of its botany, traditional uses, phytochemistry, pharmacology and toxicology

Objectives: Curcumae Radix, the medicinal part is radix, commonly called as Yujin (Chinese:), is a widely used traditional Chinese medicine for its high medicinal value and health benefits. Curcumae Radix has been used to treat conditions such as syndrome of heat Disease and unconsciousness, epilepsy and internal stagnation of phlegm, qi stagnation and blood stasis, dysmenorrhoea, jaundice, cholelithiasis caused by dampness heat of liver and gallbladder. This review aims to summarize the botany, traditional usages, processing, phytochemistry, quality control, pharmacology and toxicology of Curcumae Radix to better understand its therapeutic potential.

Key findings: So far, a variety of chemical constituents have been isolated and identified from Curcumae Radix, mainly including volatile oil and diphenylheptanes. Modern research shows that the extracts and compounds from Curcumae Radix possess wide-ranging pharmacological effects, including anti-tumour, hepatoprotective, anti-inflammatory and analgesic, AntiThrombosis, as well as effects on the nervous system and others.

Summary: Curcumae Radix holds an important position in traditional system of medicine. It is cost-effective and an important plant with curative application in contemporary medicine. However, further in-depth studies are also needed to determine the medical uses of this plant and its chemical constituents, pharmacological activity, quality control and toxicology.

The Experimental Study on Anti-thrombotic Effect of Jogantanggagambang (JGTG)

Purpose: This study was performed to evaluate anti-thrombotic effect of Jogantanggagambang extract (JGTG).

Methods: blood flow rate through the regular volume of glass tube after the blood was diluted five times with ACD solution. antithrombotic effect was calculated as a percentage of the experimental animal figure protected from the paralysis of hind legs or death of the mouse that was caused from the administration of platelet aggregation regent.

Results: 1. JGTG inhibited the platelet aggregation induced by ADP, epinephrine, collagen and arachidonic acid as compared with the control group. 2. JGTG inhibited Pulmonary Embolism induced by collagen and epinephrine (inhibitory rate is 37.5%). 3. JGTG increased platelet number and fibrinogen amount significantly and also JGTG shortened PT and APTT significantly as compared with the control group in thrombus model induced by dextran. 4. JGTG increased blood flow rate significantly as compared with the control group in vivo.

Conclusion: These results suggest that JGTG can be used for treating various female Diseases caused by Thrombosis.

Cyathula officinatis Kuan root extract

therapeutic effects of Cyathula officinalis Kuan and Its Active Fraction on Acute blood Stasis Rat Model and Identification Constituents by HPLC-QTOF/MS/MS

Objective: To evaluate the improving blood rheology and anti-inflammatory properties of C. officinalis Kuan extract (CO) and its active fraction (ACO) on acute blood stasis model Wistar rats and characterize the correlative constituents.

Materials and Methods: CO at 0.26, 0.53, and 1.04 g/kg and ACO at 0.38, 0.75, and 1.5 g/kg were administered to acute blood stasis model Wistar rats for 3 days. Whole blood viscosity, plasma viscosity, and the levels of interleukin-6 (IL-6), nitric oxide (NO), tumor necrosis factor alpha (TNF-α), and cyclooxygenase-2 (COX-2) in the plasma were measured. HPLC-QTOF/MS/MS method was used to identify the major constituents of ACO; the properties of two representative components (cyasterone and chikusetsusaponin IV) from ACO on thrombin -induced human umbilical vein endothelial cells damage model were also assessed by the levels of thromboxane A2 (TXA2), endothelin (ET), malondialdehyde (MDA), COX-2, endothelial nitric oxide synthase (eNOS), and superoxide dismutase (SOD).

Results: CO and ACO significantly reduced whole blood viscosity, plasma viscosity, and levels of IL-6, NO, TNF-α, and COX-2 in vivo. Forty compounds were identified from ACO, mainly as phytoecdysteroids and saponins. Cyasterone and chikusetsusaponin IV could significantly inhibit levels of TXA2, ET, MDA, and COX-2 and promote the activities of eNOS and SOD in vitro.

Cyathulae Radix extract

Effect of Radix Platycodonis and Radix Cyathulae in Xuefu Zhuyu Tang on tissue
distribution of paeoniflorin in blood–stasis mice by HPLC: Experimental
evidence on Shi ingredients in traditional formula compatibility

Xuefu Zhuyu Tang (XFZY), a famous formula in traditional Chinese medicine, has been demonstrated to show good therapeutic effects on Diseases caused by blood stasis syndrome. Two of its eleven herbs, Radix Platycodonis and Radix Cyathulae, have been considered as Shi ingredients in the hierarchy of traditional formula compatibility and proven possessing synergistic properties that strengthen the formula’s potency of activating blood circulation and resolving blood stagnation.

However, its mechanism is still not clearly elucidated. In our previous study, we observed their effects on paeoniflorin pharmacokinetics of XFZY in rats. In this study, we continued by detecting and comparing their effect on the tissue distribution of paeoniflorin after oral administration of XFZY and its three variants (XFZY without Radix Platycodonis or/and Radix Cyathulae) in blood-stasis mice via HPLC assay.

The results indicated that combining usage of Radix Platycodonis and Radix Cyathula increased the distribution of paeoniflorin in the lung and kidney and introduced the paeoniflorin into the liver, spleen and Heart. It might explain their synergistic properties that strengthen the formula’s effect of invigorating blood and dissolving stasis and provide experimental evidence to understand the pharmacological effects of Shi herbs in the hierarchy of traditional formula compatibility.

Virtual Screening of Potential AEC2 Inhibitors for Covid -19 from Traditional Chinese Medicine

Background: Angiotensin-converting enzyme 2 (ACE2 ), a negative regulator of the renin-angiotensin system and the severe acute respiratory syndrome-coronavirus (SARS-CoV) and SARS-CoV-2 receptor, plays an important role in viral genome replication and immune response. ACE2 has been proposed as a potential therapeutic target. Traditional Chinese medicine (TCM) could be considered as a promising complementary therapeutic option in the management of Covid -19 . However, the active components and action mechanisms that account for its therapeuti effects remain controversial.

Methods: ACE2 was employed as a target related to Covid -19 to explore the active ingredients from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) database. And the PharmMapper database and TCMSP database were used to predict the targets of the compounds. Moreover, the potential therapeutic targets of Covid -19 were acquired by intersection among genes differentially expressed in Covid -19 patients, genes screened from

Discussion of pharmacological mechanism and component compatibility of five Chinese medicines targeting ACE2 for treatment of Covid -19 using network pharmacology

Objective: To explore the action mechanism of five Chinese medicines targeting ACE2 for the treatment of Covid -19 .

Methods: Chinese medicines with human angiotensin converting enzyme II (ACE2 ) targeting activity were screened through the traditional Chinese medicine (TCM) system pharmacology database and analysis platform (TCMSP), and the TCMSP platform was further utilized to identify the active ingredients of Chinese medicines and their corresponding drug targets. Then, the protein interaction of the above protein targets was analyzed by STRING online database and the relevant data were downloaded. Next, the core sub-network modules in the PPI network system were screened through the MCODE plug-in in CytoScape 3.2.1 software. The functional enrichment analyses of gene ontology (GO) and Kyoto Encyclopedia of Gene and Genome (KEGG) were carried out, using online enrichment analysis software DAVID v6.8, to predict the pharmacological mechanism of the above five Chinese medicines, and to verify and explore the feasibility of compatibility and clinical application of related herb-pair or formula under the theoretical system of TCM.

Results: Five Chinese medicines with potential therapeutic effects on ACE2 were selected through the TCMSP platform, namely Bupleuri Radix, Puerariae Lobatae Radix, Puerariae Lobatae Flos, Cyathulae Radix, and Hemerocallis Radix, which contained 42 active ingredients and 240 drug targets. The results of GO biological function enrichment analysis showed that the above five Chinese medicines could affect the biological processes such as inflammatory response, cell apoptosis and proliferation and etc. The results of KEGG pathway enrichment analysis showed that the pharmacological mechanisms of these Chinese medicines were closely related to PI3K-Akt pathway, TNF pathway, FoxO pathway, MAPK pathway, and so on. In TCM theory, Bupleuri Radix and Puerariae Lobatae Radix is a classic herb-pair for the treatment of infectious and febrile Diseases . Chaige Jieji Decoction is widely used in febrile Diseases in ancient and modern clinics, and has achieved good clinical effects . The main febrile Disease treated by this herb-pair is the triple-yang combination of Diseases in the syndrome differentiation of the six meridians, which is in line with the syndrome differentiation of TCM in patients with SARS-CoV-2 infection. It can be used with Cyathulae Radix and Hemerocallis Radix in addition and subtraction.

Conclusion: The identified five Chinese medicines have multi-target, multi-function and multi-pathway pharmacological effects on the treatment of Covid -19 . According to the theoretical analysis of TCM and previous clinical experience, it is suggested that Chaige Jieji Decoction or Bupleuri Radix and Puerariae Lobatae Radix herb-pair can be used to treat Covid -19 .

Chinese herbal compounds against SARS-CoV-2 : Puerarin and quercetin impair the binding of viral S-protein to ACE2 receptor

The outbreak of Covid -19 raises an urgent need for the therapeutic s to contain the emerging pandemic. However, no effective treatment has been found for SARS-CoV-2 infection to date. Here, we identified puerarin (PubChem CID: 5281807), quercetin (PubChem CID: 5280343) and kaempferol (PubChem CID: 5280863) as potential compounds with binding activity to ACE2 by using Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP).

Molecular docking analysis showed that puerarin and quercetin exhibit good binding affinity to ACE2 , which was validated by surface plasmon resonance (SPR) assay. Furthermore, SPR-based competition assay revealed that puerarin and quercetin could significantly affect the binding of viral S-protein to ACE2 receptor. Notably, quercetin could also bind to the RBD domain of S-protein, suggesting not only a receptor blocking, but also a virus neutralizing effect of quercetin on SARS-CoV-2.

The results from network pharmacology and bioinformatics analysis support a view that quercetin is involved in host immunomodulation, which further renders it a promising candidate against Covid -19. Moreover, given that puerarin is already an existing drug, results from this study not only provide insight into its action mechanism, but also propose a prompt application of it on Covid -19 patients for assessing its clinical feasibility.

Dimocarpus longan Lour. extract

Fruit extract mediated synthesis of silver oxide nanoparticles using Dimocarpus longan fruit and their assesment of catalytic, antifungal, antioxidant and cytotoxic potentials

The biosynthesis of nanoparticles has been proposed as a cost-effective and environmentally friendly alternative to chemical and physical methods. In the present study, the synthesis of silver oxide nanoparticles has been achieved using Dimocarpus longan fruit. The fruit extract can act as both the reducing agent and stabilizing agent. The synthesized silver oxide nanoparticles were characterized using UV-visible spectrometer the peak observed at 460 nm, FT-IR spectrum implies that polyphenolic compounds were responsible for the formation of silver oxide nanoparticles, XRD analysis, Scanning Electron microscope (SEM) shows poly dispersed nanoparticles and EDAX spectrum shows sharp peak at 3 KeV.

The particle size range was found to be 30 to 123 nm and the stability calculated by Zeta potential was −25 mV. The prepared silver oxide nanoparticles were investigated for their catalytic activity on the reduction of p-nitrophenol(4-NP) to p-aminophenol(4-AP). The fruit extract of silver oxide nanoparticles was biologically active and exhibit antifungal activity.

The antioxidant potential of silver oxide nanoparticles were investigated by DPPH assay and the IC₅₀ value was found to be 60 μgml⁻¹. The anticancer activity of silver oxide nanoparticles was determined using MTT assay against MCF-7 breast cancer cells.

Preliminary Observation on the fibrinolytic Activity of Dimocarpus longan Seed

Dragon’s blood from Dracaena cochinchinensis (Lour.) S.C. Chen

Dragon’s blood extract has antithrombotic properties , affecting platelet aggregation functions and antiCoagulation activities

Ethnopharmacological relevance: Dragon’s blood from Dracaena cochinchinensis (Lour.) S.C. Chen (Yunnan, China), as a traditional Chinese medicinal herb, was shown to have certain antithrombotic effects.  A new preparation process was used to extract effective components from Dragon’s blood.  A 95% ethanol extract A (EA) and a precipitate B (PB) fraction were obtained and compared. Reliability of the preparation process was validated by pharmacodynamic experiments.

Materials and methods: A rat/mouse Thrombosis and blood stasis model was developed for this study, and EA and PB effects on Thrombosis, platelet functions and blood Coagulation activities were analyzed.

Results: It was observed that the EA fraction had significantly better inhibitory effects than the PB fraction on Thrombosis (p < 0.05), platelet aggregation function (p < 0.01) and anti-Coagulation activity (p < 0.05–0.01).

Conclusions: The results obtained here showed that EA fraction from Dragon’s blood contained pharmacologically effective compounds with antithrombotic effects, partially improving platelet function and antiCoagulation activity.

Comparison of the chemical profiles and anti-platelet aggregation effects of two “Dragon’s blood” drugs used in traditional Chinese medicine

Ethnopharmacological relevance: “Dragon’s blood” has been used as a medicine since ancient times by many cultures. In traditional Chinese medicine, the resin obtained from Daemonorops draco (RDD) and the resin from Dracaena cochinchinensis (RDC) are equally prescribed as “Dragon’s blood” for facilitating blood circulation.

Aim of the study: To verify the traditional efficacy and elucidate the mechanism, the present study compared the chemical profiles and the pharmacological effects of two species of “Dragon’s blood” mainly used in China.

Materials and methods: A UPLC-MS fingerprinting method was developed to compare the chemical profiles of the two medicines. The anti-platelet aggregation effects of the two medicines induced by arachidonic acid (AA) were investigated.

Results: The chemical profiles of these two species of “Dragon’s blood” were significantly different. The characteristic constituents were found to be: flavanes in RDD and stilbenes in RDC. In the in vivo platelet inhibition test, performed with the dose of 200 mg/kg on rats, the peak inhibitory effects of RDD and RDC were 35.8% and 27.6%, respectively, compared with the control group. With the in vitro concentrations of 0.2, 0.4 and 0.8 mg/ml, RDD exerted significant inhibition of aggregation by 18.7%, 20.0%, and 61.6%, respectively, and RDC exerted significant inhibition of aggregation by 13.3%, 20.2%, and 31.6%, respectively.

Conclusion: The fingerprinting method used here is suitable for distinguishing them. All pharmacological tests indicated that RDD was more potent than RDC against platelet aggregation.

Drynaria quercifolia (Polypodiaceae)

In vitro membrane stabilizing, Thrombolytic and antioxidant potentials of Drynaria quercifolia L., a remedial plant of the Garo tribal people of Bangladesh

Background: Drynaria quercifolia L. (Family- Polypodiaceae) is a fern grows in Bangladesh used in traditional healing by the Garo tribe of Mymensingh district. In the current study, rhizomes and fertile foliage fronds of this plant have been investigated comprehensively to assess their in vitro membrane stabilizing, Thrombolytic and antioxidant properties .

Methods: Rhizomes and fertile foliage fronds of D. quercifolia were collected, dried, powdered and extracted with methanol. Later on, crude methanol extracts of the plant parts were fractionated into petroleum ether, carbon tetrachloride, chloroform and aqueous soluble fractions. The extractives were then subjected to membrane stabilizing, Thrombolytic and antioxidant assays.

Results: In membrane stabilizing assay, crude methanol extracts of rhizomes and fertile foliage fronds and their petroleum ether fractions were found to be very effective for stabilizing erythrocyte membrane in hypotonic solution. In case of Thrombolytic study, crude methanol extract of rhizomes and its aqueous fraction exhibited noticeable Clotlysis . However, in antioxidant assays, crude methanol extracts of the tested plant parts and their aqueous fractions exhibited potent 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydrogen peroxide and 2, 2’-azinobis (3-ethylbenzothiazoline sulphonic acid) (ABTS) radical scavenging activity. Besides, these extractives also displayed substantial ferric reducing potential in ferric reducing antioxidant power (FRAP) assay. Crude methanol extracts of the plant parts and their aqueous fractions were also found rich in phenolics.

Conclusion: This study demonstrates the medicinal potentials of D. quercifolia and justifies the local uses of it by the Garo tribal people of Bangladesh for multiple Disease management.

CLINICAL APPRAISALS AND PHYTOCHEMICAL POTENTIAL OF ETHNOMEDICINAL PTERIDOPHYTE: DRYNARIA QUERCIFOLIA (L.) J. SMITH (POLYPODIACEAE)

Drynaria quercifolia is a medicinal pteridophyte which have been treated by the administration of plant parts based on traditional and folk uses since ancient times. Considerable utilization and progress have been achieved regarding its biological activities. Various phytoconstituents like 3,4-dihydroxybenzoic acid, friedelin, epifriedelinol, coumarins β-amyrin, β-sitosterol and β-sitosterol 3-β-D-glucopyranoside has been isolated from the plant and these bioactive compounds responsible for its antidermatophytic, antimicrobial, antifertility, anti-lipidperoxidative, antiulcer, antipyretic, anti-arthritis, anti-urolithiatic, Pesticidal and Pest Repellency, Thrombolytic and various other activities.

Considering the ethnomedicinal significance of this parasitic fern, an attempt has been made for the first time to review to provide up to date clinical reports on the plant and to document the available phytochemical constituents through data base searches (PubMed, Google scholar, Scopus).

EGb 761 from Ginkgo biloba extract

Antiplatelet and antithrombotic effects of a Combination of Ticlopidine and Ginkgo biloba Ext (EGb 761)

The antiplatelet and antithrombotic effects of the oral combination treatment of ticlopidine and Ginkgo biloba extract (EGb 761) were studied in normal and Thrombosis -induced rats. The ex vivo inhibitory effect on ADP-induced platelet aggregation of a small dose of ticlopidine (50 mg/kg/day) in combination with EGb 761 (40 mg/kg/day) was comparable to a larger dose of only ticlopidine (200 mg/kg/day). Bleeding time was also prolonged by 150%. thrombus weight was also consistently decreased by a combination of ticlopidine and EGb 761 in an arterio-Venous shunt model at two doses of ticlopidine (50 mg/kg) plus EGb 761 (20 mg/kg) and ticlopidine (50 mg/kg) plus EGb 761 (40 mg/kg). A combinatory treatment in acute Thrombosis model in mice also showed a higher recovery than a single treatment.

effects of Ginkgo biloba extract (EGb 761) on cerebral Thrombosis and blood pressure in Stroke -prone spontaneously hypertensive rats

1. An extract of Ginkgo biloba (EGb 761) has been reported to alleviate cerebrovascular problems. In the present study, we investigated the antithrombotic effects of EGb 761 in cerebral blood vessels of Stroke -prone spontaneously hypertensive rats (SHRSP/Izm).

2. In the present study, EGb 761 was administered orally to SHRSP/Izm at 60 and 120 mg/kg each day for 3 weeks from the age of 7 weeks. The age-related increase in blood pressure observed in SHRSP was suppressed significantly by EGb 761 at both doses 3 weeks after treatment.

3. Thrombotic potential was assessed in vivo using a He-Ne laser-induced Thrombosis model and was significantly suppressed by EGb 761.

4. The anti-oxidant effects of EGb 761 were determined by measurement of urinary 8-hydroxy-2′-deoxyguanosine (8-OHdG). At 120 mg/kg, EGb 761 decreased 8-OHdG significantly compared with control animals.

5. Urinary nitrite/nitrate, nitric oxide (NO) metabolites, were increased significantly after administration of EGb 761. Expression of endothelial NO synthase (eNOS) mRNA was measured using a real-time quantitative reverse transcription–polymerase chain reaction method. The expression of eNOS mRNA in the EGb 761 group (120 mg/kg) was significantly higher than in the control group.

6. The results indicate that EGb 761 decreases blood pressure and mediates strong antithrombotic and anti-oxidant effects in SHRSP. These pharmacological activities may contribute to the beneficial properties of EGb 761 observed in clinical practice.

fibrinolytic effects of Ginkgo biloba extract

A multitude of factors are involved in regulating the blood Coagulation homeostatic processes in the body, which may ultimately lead to thromboemboli and Thrombosis. The resolution of blood clots after healing is as important as Clot formation at the site of a vascular lesion. This is accomplished by fibrinolytic drugs such as streptokinase (SK) and urokinase.

It must be noted that administration of SK may be accompanied by the lysis of blood Clot s in unwanted sites, and complications such as general lytic conditions, severe hemorrhaging, reduced serum fibrinogen and allergies can occur. Anti-SK antibodies neutralize the effects of SK. Studies on natural compounds and medicinal herbs with fewer side effects have been ongoing. In the present study, the fibrinolytic effect of Ginkgo biloba, an herb grown in Iran, was investigated.

A polyphenolic method was used to obtain Ginkgo extract from its leaves. The fibrinolytic effects of SK (positive control) were compared with those of Ginkgo extract using a fluorometry method.

In producing a labelled Clot, fibrinogen was labelled with the fluorescent agent fluorescein isothiocyanate and precipitated in the presence of Ca2+. SK (100 U/mL to 1000 U/mL) and Ginkgo extract were added to labelled fibrin in a plasma environment at dilutions of 1, 1:10, 1:100 and 1:1000 (volume/volume). The fluorescence of the solution was measured between 15 min and 60 min later.

A linear relationship was observed between the fluorescence measured and SK concentrations ranging from 300 U/mL to 700 U/mL. Ginkgo extract displayed a remarkable effect in resolving the Clot. As Ginkgo extract remained in the environment, fluorescence increased notably, showing a time-dependent relationship.

Overall, the results indicate that the effects of Ginkgo extract on the fibrinolytic system are similar to those of SK; hence, this herbal extract can be used as a complement to or a substitute for SK. Additionally, it is proposed that the effects of the active ingredients of Ginkgo extract should be studied in animals. Further studies are warranted for evaluating the possible side effects and toxicity of Ginkgo extract in human subjects.

Inhibitory effect of Ginkgo biloba extract on human platelet aggregation

The effect of pure flavonoids and Gingko biloba extract (GBE) on human platelet aggregation was investigated. Most of the flavonoids and vitamin E did not affect platelet aggregation in platelet-rich plasma (PRP); however some of these flavonoids inhibited platelet aggregation in gel-filtered platelets (GFP). GBE inhibited both ADP- and collagen-induced platelet aggregation in PRP, GFP and in whole blood in a dose-dependent manner. GBE at very low concentrations inhibited whole blood aggregation induced by ADP compared with those used for PRP or GFP. Flavonoids and GBE decreased the production of TxA 2 induced by collagen and ADP in PRP.

owever, no correlation was observed between the inhibition of platelet aggregation and the decrease of TxA 2 synthesis. GBE and flavonoids did not affect platelet membrane fluidity. However, the incubation of PRP with GBE increased cAMP levels in platelets, which is known to inhibit platelet activation by lowering intracellular Ca2+ levels. GBE is a mixture of many compounds, including flavonoids and gingkoglides, which affect metabolism of cAMP, TxA2 and Ca2+ in platelets. It is effective in the inhibition of platelet aggregation, both in PRP and whole blood, and thus may be potentially used as an effective oral anti-platelet therapeutic agent.

effects of Ginkgo biloba on blood Coagulation Parameters

A systematic literature review was conducted to investigate the effects of Ginkgo biloba preparations on blood Coagulation parameters. Systematic literature searches were carried out using MEDLINE (via PubMed), EMBASE, AMED, CINAHL®, the Cochrane Library and PsycINFO to identify randomised, double-blind, clinical trials of G. biloba mono-preparations assessing at least one blood Coagulation parameter as either a primary or a supplementary outcome measure. Five manufacturers of G. biloba preprations were asked to contribute any unpublished material. Data were validated and extracted independently by two reviewers according to pre-specified criteria.

Eight trials were included, of which two included healthy subjects, four included patients with various conditions and two included participants taking concomitant Anticoagulant medication, one with acetylsalicylic acid (ASA; aspirin) and one with warfarin. Overall results do not provide evidence that G. biloba does affect blood Coagulation parameters in a clinically relevant manner. Out of many parameters assessed, four Coagulation parameters from three studies were significantly different in the treatment group compared with the control. One study comparing the treatments of ASA and G. biloba with ASA alone confirmed that the two treatments were equivalent with regards to all but three Coagulation parameters.

Available evidence does not demonstrate that extract of G. biloba causes significant changes in blood Coagulation parameters. A limited number of studies on concomitant use of G. biloba with ASA or warfarin also do not suggest that G. biloba has an additive effect to the clinical effects of these two drugs. Currently available evidence is limited and, therefore, further studies are needed to assess effects of G. biloba on a variety of blood Coagulation parameters.

Evodia rutaecarpa (Juss.) Benth extract

antithrombotic effect of rutaecarpine, an alkaloid isolated from Evodia rutaecarpa, on platelet plug formation in in vivo experiments

In this study, platelet thrombi formation was induced by irradiation of mesenteric venules with filtered light in mice pretreated intraVenous ly with fluorescein sodium. Rutaecarpine (200 µg/g) significantly prolonged the latent period of inducing platelet plug formation in mesenteric venules when it was intravenously injected. Rutaecarpine (200 µg/g) prolonged occlusion time by approximately 1·5-fold (control 127 ± 29 vs. taecarpine 188 ± 23 s). Furthermore, aspirin (250 µg/g) also showed a similar prolongation of the occlusion time in this experiment. On a molar basis, rutaecarpine was approximately twofold more potent than aspirin at prolonging the occlusion time.

Furthermore, rutaecarpine was also effective in reducing the mortality of ADP-induced acute Pulmonary thromboEmbolism in mice when administered intravenously at doses of 25 and 50 µg/g. Intravenous injection of rutaecarpine (50 µg/g) significantly prolonged the bleeding time by approximately 1·5-fold compared with normal saline in the severed mesenteric arteries of rats. Continuous infusion of rutaecarpine (5 µg/g/min) also significantly increased the bleeding time 1·5-fold, and the bleeding time returned to baseline within 60 min after cessation of rutaecarpine infusion.

These results suggest that rutaecarpine has an effective anti-platelet effect in vivo and that it may be a potential therapeutic agent for Arterial Thrombosis, but it must be assessed further for toxicity.

Rutaecarpine, an Alkaloid from Evodia rutaecarpa, Can Prevent platelet activation in Humans and Reduce Microvascular Thrombosis in Mice: Crucial Role of the PI3K/Akt/GSK3β Signal Axis through a Cyclic Nucleotides/VASP—Independent Mechanism

The role of activated platelet s in acute and chronic Cardiovascular Diseases (CVDs) is well established. Therefore, antiplatelet drugs significantly reduce the risk of severe CVDs. Evodia rutaecarpa (Wu-Chu-Yu) is a well-known Chinese medicine, and rutaecarpine (Rut) is a main bioactive component with substantial beneficial properties including vasodilation . To address a research gap, we investigated the inhibitory mechanisms of Rut in washed human platelets and experimental mice. At low concentrations (1–5 μM), Rut strongly inhibited collagen-induced platelet aggregation, whereas it exerted only a slight or no effect on platelets stimulated with other agonists (e.g., thrombin ).

Rut markedly inhibited P-selectin expression; adenosine triphosphate release; [Ca2+]i mobilization; hydroxyl radical formation; and phospholipase C (PLC)γ2/protein kinase C (PKC), mitogen-activated protein kinase, and phosphoinositide 3-kinase (PI3K)/Akt/glycogen synthase kinase-3β (GSK3β) phosphorylation stimulated by collagen. SQ22536 (an adenylate cyclase inhibitor) or ODQ (a guanylate cyclase inhibitor) did not reverse Rut-mediated antiplatelet aggregation. Rut was not directly responding to vasodilator-stimulated phosphoprotein phosphorylation.

Rut significantly increased the occlusion time of fluorescence irradiated thrombotic platelet plug formation. The findings demonstrated that Rut exerts a strong effect against platelet activation through the PLCγ2/PKC and PI3K/Akt/GSK3β pathways. Thus, Rut can be a potential therapeutic agent for Thromboembolic disorders.

Effect of Evodiamine on Collagen-Induced platelet activation and Thrombosis

Evodia rutaecarpa has multiple pharmacological effects and is widely used in the prevention and treatment of migraine, diabetes, Cardiovascular Disease, cancer, and other chronic Diseases; however, the pharmacological effects of its active compound evodiamine (Evo) have not been thoroughly investigated. The purpose of this study was to investigate the effects of Evo on antiplatelet activation and Thrombosis. We discovered that Evo effectively inhibited collagen-induced platelet activation but had no effect on platelet aggregation caused by activators such as thrombin, ADP, and U46619. Second, we found that Evo effectively inhibited the release of platelet granules induced by collagen.

Finally, evodiamine inhibits the transduction of the SFKs/Syk/Akt/PLCγ2 activation pathway in platelet s. According to in vivo studies, Evo significantly prolonged the mesenteric thrombo embolism induced by ferric chloride and had no discernible effect on the Coagulation function of mice. In conclusion, the antiplatelet and thrombotic effects of Evo discovered in this study provide an experimental basis for the investigation of the pharmacological mechanisms of Evo and the development of antiplatelet drugs.

Cardiovascular Pharmacological Actions of Rutaecarpine, a Quinazolinocarboline Alkaloid Isolated From Evodia rutaecarpa

Evodia rutaecarpa (Chinese name: Wu-Chu-Yu) is a well-known traditional Chinese medicine and has long been used in Chinese medical practice. Rutaecarpine is an indolopyridoquinazolinone alkaloid isolated from E. rutaecarpa and related herbs, which has been shown to have Cardiovascular biological effects, such as inotropic and chronotropic, vasorelaxant, antiplatelet aggregation, and anti-inflammatory effects.

Furthermore, it has been reported that rutaecarpine has beneficial effects on some Cardiovascular Diseases. This review was undertaken to summarize data on the Cardiovascular pharmacological actions of rutaecarpine published over the recent years, aiming to provide more evidence supporting its use in the treatment of Cardiovascular Diseases. This review also reveals some interesting and unique pharmacological properties, which may explain its vascular and platelet effects.

Rutaecarpine: A promising Cardiovascular protective alkaloid from Evodia rutaecarpa (Wu Zhu Yu)

Rutaecarpine is a bioactive alkaloid isolated from Evodia rutaecarpa (Wu Zhu Yu, Family: Rutaceae), a versatile medicinal herb which is clinically used to treat headache, abdominal pain, postpartum hemorrhage, dysentery, and amenorrhea in China. As one of the most representative indolopyridoquinazoline alkaloids of Evodia rutaecarpa, rutaecarpine has broad pharmacological actions in treating various Cardiovascular, cerebrovascular, and metabolic diseases.

The Cardiovascular actions of rutaecarpine have aroused intense research interest due to its purported inotropic and chronotropic, vasodilatory, anti-platelet activation, anti-oxidant, anti-inflammatory, and lipid-lowering effects. Biochemical and pharmacological studies have illustrated the molecular targets of rutaecarpine, such as TRPV1, CGRP, AMPK, ABCA1, and β1-AR. Furthermore, several rutaecarpine derivatives (such as bromorutaecarpine and fluororutaecarpine) have been shown to possess cardioprotective and vasculoprotective effects with improved safety profile.

Hereby, we provide a systematic overview of pharmacological actions, toxicological effects, and molecular targets of rutaecarpine in Cardiovascular Disease prevention/treatment, aiming to exploit the therapeutic potential of rutaecarpine and its derivatives in treating Cardiovascular Diseases.

Ganoderma lucidum

fibrinolytic and antithrombotic protease from Ganoderma lucidum

A putative metalloprotease was purified from mycelium of Ganoderma lucidum. The enzyme was purified by anion exchange, chromatofocusing, and gel filtration chromatography. The Mr was determined to be 52 000 by SDS-PAGE and 100 000 by gel filtration on a Sephadex G-150 column, indicating that it is a dimer. The enzyme was inhibited by EDTA, 1,10-phenanthroline, and phosphoamidon. The presence of Zn2+ was detected by ICP mass spectral analysis as 1.1 mol of Zn2+ per mol of protease.

This protease hydrolyzed Aα and Bβ chains of human fibrinogen, but did not cleave thrombin, albumin, hemoglobin, and immunoglobulin under the same condition. It also showed an Anticoagulant activity in human plasma. The enzyme delayed the activated partial thromboplastin time and thrombin time, but not the Clot ting induced by reptilase, indicating Ganoderma protease is specific to thrombin. Ganoderma protease behaved as a competitive inhibitor of thrombin-catalyzed fibrin formation, but no inhibition of thrombin was found with a small synthetic peptide. These observations indicate that Ganoderma protease could bind thrombin at an anion binding exosite distinct from the active site and cause the delay of Clot ting time induced by thrombin.

Studies on Screening, Isolation and Purification of a fibrinolytic Protease from an Isolate (VK12) of Ganoderma Lucidum and Evaluation of its antithrombotic Activity

antithrombotic activity of a protease purified from a medicinal mushroom, Ganoderma lucidum, has been evaluated platelet aggregation in vitro and Pulmonary Thrombosis in vivo. The purified protease exhibited concentration dependent inhibitory effects on platelet aggregation induced by ADP (adenosine diphosphate), with an IC₅₀ value of 2.4 mg/mL. The purified protease protected mice against thrombotic death or paralysis induced by collagen and epinephrine in a dose dependent manner when administered orally. It produced a significant inhibition of thrombotic death or paralysis at 60μg/kg body weight, while aspirin produced a significant inhibition of Thrombosis at 10-20 mg/kg body weight.

The purified protease also has showed fibrinolytic activity and alters Coagulation parameters such as activated partial thromboplastin time (APTT), and thrombin time (TT) in rat platelet. These results suggested that the antithrombotic activity of Ganoderma lucidum protease might be due to antiplatelet activity rather than antiCoagulation activity.

In vitro Doses and Incubations Dependent Thrombolytic Potential Study of Edible Mushrooms Pleurotus ostreatus, Ganoderma lucidum and Lentinula edodes Available in Bangladesh

Aims: The study was carried out to investigate the Thrombolytic potential of methanolic extracts of edible mushrooms.

Study Design: Experimental study. Place and Duration of Study: Department of Biochemistry, Primeasia University, August 2014 to December 2014

Methodology: An in vitro Thrombolytic model was used to check the Thrombolytic effect of Pleurotus ostreatus, Ganoderma lucidum and Lentinula edodes along with Streptokinase as a positive control and water as a negative control. In this study two concentrations of the extracts i.e. 400μg/ml and 800 μg/ml were tested at three time intervals, 24 hrs, 48 hrs and 72 hrs duration of incubation at 37ºC for observing maximum Clotlysis.

Results: At 400 μg/ml dose, Pleurotus ostreatus showed highest Clotlysis activity in 72 hrs of incubation, 18.62% while Ganoderma lucidum, and Lentinula edodes showed 17.01% and 9.02% in the same incubation. Similarly, at 800 μg/ml dose, the Thrombolytic activities were 63.35%,

Gastrodiae Rhizoma extract

effects of Acidic Polysaccharides from Gastrodia Rhizome on Systolic blood Pressure and Serum Lipid Concentrations in Spontaneously Hypertensive Rats Fed a High-Fat Diet

The effects of acidic polysaccharides purified from Gastrodia rhizome on blood pressure and serum lipid levels in spontaneously hypertensive rats (SHR) fed a high-fat diet were investigated. Acidic polysaccharides were purified from crude polysaccharides by DEAE-Sepharose CL-6B. Thirty-six male SHR were randomly divided into three groups: Gastrodia rhizome crude polysaccharide (A), acidic polysaccharide (B) groups, and a control group (C). A 5-week oral administration of all treatment groups was performed daily in 3- to 8-week-old SHRs with a dose of 6 mg/kg of body weight/day.

After 5 weeks of treatment, total cholesterol in the acidic polysaccharide group, at 69.7 ± 10.6 mg/dL, was lower than in the crude polysaccharide group (75.0 ± 6.0 mg/dL) and the control group (89.2 ± 7.4 mg/dL). In addition, triglyceride and low-density lipoprotein cholesterol levels in the acidic polysaccharide group were lower than in the crude polysaccharide and control groups. The atherogenic index of the acidic polysaccharide group was 46.3% lower than in the control group.

Initial blood pressure after the initial three weeks on the high-fat diet averaged 195.9 ± 3.3 mmHg among all rats. Compared with the initial blood pressure, the final blood pressure in the control group was increased by 22.8 mmHg, whereas it decreased in the acidic polysaccharide group by 14.9 mmHg. These results indicate that acidic polysaccharides from Gastrodia rhizome reduce hypertension and improve serum lipid levels.

Gastrodia elata Blume Rhizome Aqueous Extract Improves Arterial Thrombosis , Dyslipidemia, and Insulin Response in Testosterone-Deficient Rats

Testosterone deficiency deteriorates glucose and lipid metabolism with reducing muscle mass. We investigated whether the consumption of water extracts of Gastrodia elata Blume rhizome (GEB) rich in gastrodin would reduce the symptoms of testosterone deficiency and improve blood flow in orchidectomized (ORX) rats. ORX rats were given high-fat diets supplemented with either 1% cellulose (ORX-control), 0.3% GEB (GEB-L), or 1% GEB (GEB-H) for 8 weeks.

Sham-operated rats were fed the same diet as OVX-control rats (normal-control). ORX-control rats had reduced serum testosterone levels by one-fifth, compared to normal-control rats. ORX-control rats exhibited decreased lean body mass, attenuated blood flow, and impaired cholesterol metabolism and glucose control due to decreased insulin secretory response. GEB increased serum insulin levels dose-dependently and GEB-H mostly enhanced dyslipidemia in ORX rats. GEB completely normalized Arterial Thrombosis time and blood flow in ORX rats.

Interestingly, ORX-control rats showed attenuated hepatic insulin signaling but greater AMPK and CREB activities, which reduced triglyceride accumulation, compared to normal-control. GEB-H improved hepatic insulin signaling but maintained the AMPK and CREB activities in ORX rats. In conclusions, GEB ameliorated the impairment of cholesterol and glucose metabolism and blood flow in ORX rats. GEB may be a potential preventive measure for reducing the risk of Cardiovascular Diseases associated with testosterone deficiency.

Haliotis diversicolor Reeve extract

In vitro anti-thrombotic and anti-coagulant properties of blacklip abalone (Haliotis rubra) viscera hydrolysate

Abalone viscera contain sulphated polysaccharides with anti-thrombotic and anti-coagulant activities. In this study, a hydrolysate was prepared from blacklip abalone (Haliotis rubra) viscera using papain and bromelain and fractionated using ion exchange and size exclusion chromatography. Hydrolysates and fractions were investigated for in vitro thrombin inhibition mediated through heparin cofactor II (HCII) as well as anti-coagulant activity in plasma and whole blood. On the basis of sulphated polysaccharide concentration, the hydrolysate inhibited thrombin through HCII with an inhibitor concentration at 50% (IC50) of 16.5 μg/mL compared with 2.1 μg/mL for standard heparin.

Fractionation concentrated HCII-mediated thrombin inhibition down to an IC50 of 1.8 μg/mL and improved anti-coagulant activities by significantly delaying Clot ting time. This study confirmed the presence of anti-thrombotic and anti-coagulant molecules in blacklip abalone viscera and demonstrated that these activities can be enriched with a simple chromatography regime. Blacklip abalone viscera warrant further investigation as a source of nutraceutical or functional food ingredients.

Anti-coagulant and Anti-Thrombotic properties of Blacklip Abalone (Haliotis rubra): In Vitro and Animal Studies

Sulphated polysaccharides with anti-thrombotic and anti-coagulant activities have been found in various marine biota. In this study, a previously characterised anti-thrombotic and anti-coagulant extract from blacklip abalone was fractionated by anion exchange chromatography (AEC), pooled (on a sulphated polysaccharide basis) and administered to Wistar rats via oral gavage (N = 8) for assessment as an oral therapeutic.

To ensure that the preparation had anti-coagulant activity prior to oral administration, it was assessed in rat blood by thromboelastography (TEG) significantly increasing reaction (R) time (or time until Clot formation). Following in vitro confirmation of anti-coagulant activity, 40 mg of the preparation was orally administered to rats with blood samples collected at 2, 4, and 6 h post-gavage. Assessment of all blood samples by TEG showed some prolongation of R time from 355 to 380 s after 4 h. Dosing of the post-gavage blood samples with the abalone preparation to confirm anti-thrombotic activity in vitro revealed residual anti-coagulant activity, further suggesting that oral administration did increase anti-coagulant potential in the collected blood but that bioavailability was low.

Assessment of tissues and haematological parameters showed no obvious harmful effects of the abalone preparation in animals. In summary, even though oral administration of fractionated and pooled blacklip abalone extract to rats delayed clotting after 4 h, bioavailability of the preparation appeared to be low and may be more appropriate for intravenous administration as an anti-thrombotic or anti-coagulant therapeutic.

antithrombotic potential of extracts from abalone, Haliotis Discus Hannai Ino: In vitro and animal studies

To screen the more promising antithrombotic abalone extracts, the in vitro antithrombotic activity of 6 abalone (Haliotis Discus Hannai Ino) extracts was examined in this study. Results of in vitro studies indicated that water extract of abalone viscera (WEV) presented significant longer prothrombin time (PT), partial thromboplasmatin time (APTT), and thrombin time (TT) than other abalone extracts. In addition, the platelet aggregation process was inhibited by WEV as well. The antithrombotic potential of WEV was further investigated in animal studies which revealed that WEV prolonged tail bleeding time, APTT but not PT and TT and showed no effect on the platelet aggregation and platelet number. The WEV treated rats presented lower cholesterol and triglyceride level than the control group although the difference was not statistically significant.

Results of this study indicate the antithrombotic activity of abalone extracts, especially WEV, and may contribute to the further study in this field.

Hawthorn berry extract

Handbook of nutrition in Heart health

The growing body of evidence suggests that aggressive treatment for the risk factor management will improve survival and reduce the Cardiac events. It is well known Mediterranean diets enriched in plant foods are associated with positive health outcomes in reducing risk factors of Cardiovascular Disease (CVD). This review summarizes plant bioactive nutrients such as resistant starch, omega-3 fatty acids source from walnuts, avocados, coenzyme Q10, hawthorn berry fruit extract, curcumin/curcuminoids, lycopene, gugulipid and gotu kala plant sources impact on cardio-metabolic risk factors.

These plant sources of bioactive nutrients exhibited protective effects against hyperglycemia, hyperlipidemia, inflammation and oxidative stress, known risk factors for CVD and diabetic complications, in cell culture, animal studies and some human studies. This review provides information on plant bioactive nutrients, with a particular focus on active nutrients involved in lipid, carbohydrate and metabolic pathways and their potential health benefits in relation to the prevention and treatment of risk factors for hypertension, endothelial function, insulin resistance, type 2 diabetes, weight management, microalbuminuria, hypercoagulability, inflammation, abnormal thrombolysis, increased oxidative stress and CVD.

antithrombotic effects of ethanol extract of Crataegus orientalis in the carrageenan-induced mice tail Thrombosis model

Materials and Methods: Antithrombotic function of ethanol extract of Crataegus orientalis (COE) leaves was investigated in carrageenan-induced mice tail Thrombosis model. Mice were injected with 40 μl (1%) carrageenan (Type I) dissolved in physiological saline by intraplantar administration in the right hind paw. After carrageenan injection, the extract was administered at the doses of 100, 200, and 300 mg/kg. Heparin was used as a positive control (10 and 100 IU). The length of tail-Thrombosis was measured at 24th, 48th, and 72nd hours.

Evaluation of the antithrombotic effects of Crataegus monogyna and Crataegus davisii in the carrageenan-induced tail Thrombosis model

Context: Crataegus species are widely used as herbal medicines for preventing Cardiovascular Diseases (CVDs). However, there are no studies on the effects of Crataegus monogyna Jacq. (Rosaceae) and C. davisii Browicz on Thrombosis, which is an important mechanism in CVDs.

Objective: The purpose of this study was to investigate the antithrombotic effects of ethanol extracts of Crataegus monogyna (CMEx) and C. davisii (CDEx) leaves by using the carrageenan-induced tail Thrombosis model.

Materials and methods: The hind paw of each mouse was injected with 1% Type I carrageenan to induce Thrombosis. CMEx was tested at the doses of 100, 200, and 300 mg/kg and CDEx at the dose of 50, 100, 200, and 300 mg/kg in comparison with heparin. The lengths of tail Thrombosis were measured at the 24, 48, and 72 h.

Results: Does of 200 and 300 mg/kg CMEx showed significant effects (p < 0.01; p < 0.001) at 24 h when compared with the control group. The antithrombotic activity of 200 and 300 mg/kg CMEx showed a decrease at 48 and 72 h but the activity of 300 mg/kg dose of CMEx was still significant (p < 0.01). The activities of 50 and 100 mg/kg doses of CDEx were significant (p < 0.001; p < 0.01) between 24 and 72 h whereas 200 and 300 mg/kg CDEx did not show any significance.

Discussion and conclusions: CMEx and CDEx significantly inhibited the carrageenan-induced mouse tail Thrombosis. Based on these results, it was concluded that CDEx and CMEx may potentially be used as therapeutic agents or complementary treatments against Thrombosis.

AN OVERVIEW: NATURAL HERBS AS AN ATHERO-ThrombolyticS

Atherosclerosis is the preliminary lipid disorder in which Arterial blood vessels become hard and thicken due to multiple Plaque formation within blood vessels. These soft multiple Plaques suddenly ruptures out, leading to the formation of thrombus that rapidly slow down or stop the blood flow and obstruct the oxygen supply to the targeted tissues. These free floating thrombi can be circulated anywhere in the Cardiovascular system including lung, brain, Heart or Deep veins of leg and leads to serious complications as atherothrombotic Diseases (Myocardial or cerebral Infarction), Coronary Artery Diseases (CAD), peripheral vascular Diseases, Pulmonary Thrombosis, cerebro-vascular Disease and Heart failure.

Pulmonary Thrombosis is a inexorable condition commonly appear in Covid -19 pneumonia patients due to fully or partially block blood supply to Artery of lung and is characterized by inexplicable breathing difficulties, hemoptysis and chest pain. Similarly cardiac arrhythmias is on rise worldwide and is leading cause of morbidity, mortality and disabilities, is associated with shortness of breath, synapse, angina or finally even death. A Cardio vascular Disease (CVD) is the highly implacable Disease and often impacting most productive and precious years of individuals throughout the world and hence requires very accurate diagnosis and treatment. Commonly used conventional antithrombotic drugs such as t-PA, Urokinase, Streptokinase, Ateplase, Anisteplase and so forth, play deciding role in CVD management and other related disorders.

However, available Thrombolytics still have noticeable shortcomings including bleeding, lysis of hemolytic plug, antigenicity, defective Coagulation , gastrointestinal and cerebral hemorrhage. In view of the inadequacies of conventional Thrombolytic s, efforts have made to understand significant value of herbal drugs for treating Coronary Arterial Disease and related problems. As diabetes mellitus, hypertension, smoking cigarettes, dislipidemia, dyspnea and pneumonia are the prime causes of increasing severity of atherosclerosis and Covid -19 Diseases , includes aggregation of platelets which eventually results in thrombus formation and hypoxia.

Moreover, Covid – 19 death rate is more common among people with such Diseases and disorders. Hence food and dietary herbals with antithrombotic characteristics may be used to improve such Diseases which ultimately maintain the health and regain healthy state of mind. The present review is focused on the availability of medicinal plants that have antithrombotic , anti- platelets and fibrinolytic potential, which can be explored for the effective treatment of thrombotic Diseases , lifestyle disorders and future pandemics like Covid -19.

kaempferol

Kaempferol inhibits Thrombosis and platelet activation

The objectives of the present study were to investigate whether kaempferol affects pro-coagulant proteinase activity, fibrin Clot formation, blood Clot and thrombin (or collagen/epinephrine)-stimulated platelet activation, Thrombosis, and Coagulation in ICR (Imprinting Control Region) mice and SD (Sprague–Dawley) rats. Kaempferol significantly inhibited the enzymatic activities of thrombin and FXa by 68 ± 1.6% and 52 ± 2.4%, respectively. Kaempferol also inhibited fibrin polymer formation in turbidity. Microscopic analysis was performed using a fluorescent conjugate. Kaempferol completely attenuated phosphorylation of extracellular signal-regulated kinase (ERK) 1/2, p38, c-Jun N-terminal kinase (JNK) 1/2, and phosphoinositide 3-kinase (PI3K)/PKB (AKT) in thrombin-stimulated platelets and delayed aggregation time (Clot ting) by 34.6% in an assay of collagen/epinephrine-stimulated platelet activation.

Moreover, kaempferol protected against Thrombosis development in 3 animal models, including collagen/epinephrine- and thrombin-induced acute thromboEmbolism models and an FeCl3-induced carotid Arterial thrombus model. The ex vivo Anticoagulant effect of kaempferol was further confirmed in ICR mice. This study demonstrated that kaempferol may be clinically useful due to its ability to reduce or prevent thrombotic challenge.

lactoferrin

In this study, researchers at Soochow University in China investigated the effects of lactoferrin (LF) supplementation on atherosclerosis. They reported their findings in an article published in the Journal of Medicinal Food.

LF is a multifunctional glycoprotein in milk with beneficial effects on lipid metabolism. However, whether LF can inhibit the development of atherosclerosis is yet to be explored. The researchers fed 48 male apolipoprotein E?/? mice with a high-fat diet that contains 1.25 percent added cholesterol. They divided the mice into four treatment groups: the group given distilled water (HFCD), the group given 2 mg/mL LF solution, the group given 10?mg/mL LF (MLF) and the group given 20?mg/mL LF (HLF) for 12 weeks. The researchers conducted glucose tolerance tests (OGTT) at weeks 0, 4, 8 and 12 and measured serum, liver and fecal lipid levels at the end of the experiment.

They also conducted a pathological examination of the livers, whole aortas and aortic sinuses of the mice and measured the expression of proteins related to cholesterol synthesis, absorption and excretion. The researchers reported no significant difference between the body weight, food intake and OGTT results of the four groups. Compared with the HFCD group, the MLP and HLF groups had significantly reduced serum and liver cholesterol levels but significantly increased fecal cholesterol content. Treatment with LF alleviated hepatic steatosis and lipid droplet formation, as evident in the MLF group.

LF also significantly decreased the average lesion areas in the whole aorta, especially in the MLF group. LF decreased the expression of HMG-CoA reductase — a rate-limiting enzyme involved in cholesterol synthesis — in the liver but increased the expression of cholesterol 7-alpha hydroxylase, a rate-limiting enzyme involved in the synthesis of bile acid from cholesterol. LF also decreased the expression of Niemann-Pick C1-like 1 protein, which binds to a critical mediator of cholesterol absorption, in the intestines. Based on these findings, the researchers concluded that lactoferrin alleviates atherosclerosis by decreasing cholesterol synthesis and absorption and increasing cholesterol excretion.

Lagerstroemia speciosa

Thrombolytic activity of Lagerstroemia speciosa Leaves

Aim: to investigate the Thrombolytic activity of Lagerstroemia speciosa leaf extract.

Methods: Thrombolytic activity assayed by an in vitro Thrombolytic  activity model performed on 10 apparently healthy subjects (both male and female), where the Clotlysis ability of methanolic extract of Lagerstroemia speciosa leaves compared with Streptokinase a positive control and saline water as a negative control.

Result: Fraction of methanolic extract of Lagerstroemia speciosa exhibited significant percentage (%) of Clotlysis of 25.42% compared to positive control streptokinase of 31.06% loss of Clo while the negative control saline water showed 3.81% Clotlysis ability.

Conclusion: Thepresentinvestigationrevealedthatthemethanolextracts of the leaves of Lagerstroemia speciosa possess potent Thrombolytic properties may refer to the treatment of Cardiovascular Diseases.

antithrombotic and antihemolytic effects of Lagerstroemia speciosa (Lythraceae) aqueous extract

Introduction: Aqueous extract of Lagerstroemia speciosa (EALS) (Lythraceae) is widely used to treat diabetes. This plant has been shown an in vitro Thrombolytic activity that indicates its potential to prevent the formation of blood Clot s in vivo. Thus, this study was undertaken to evaluate the antithrombotic and antihemolytic effects of EALS.

Methods: Rats of both sexes (200 ± 5 g) were divided into five groups of six animals. Each group received orally distilled water, EALS (250, 500, 1000 mg/kg), and acetylsalicylic acid (100 mg/kg) for five days. After treatment, the FeCl3-induced Arterial thrombus formation method was used to determine occlusion time. A coagulometer was used to detect activated partial thromboplastin time (aPTT) and prothrombin time (PT). Rabbit blood was used to determine Clotlysis activity in vitro and antihemolytic activity using the 2,2-azobis hydrochloride (2-methylpropionamidine) (AAPH) method.

Results: EALS increased the occlusion time in a dose-dependent manner. At the dose of 1000 mg/kg, EALS increased the occlusion time significantly, from 4.59 ± 2.45 minutes to 15.52 ± 2.38 minutes (P < 0.01). At high concentrations (1-4 mg/mL), EALS showed a significant increase in aPPT and PT (P < 0.05). Streptokinase and EALS (4 mg/mL) induced significant Clotlysis with percentage values of 78.48 ± 2.2 % and 49.5 ± 1.53 %, respectively (P < 0.001). EALS inhibited AAPH-induced hemolysis.

Conclusion: EALS exhibited antithrombotic and antihemolytic activities. The antithrombotic property of the plant could be attributed to its Anticoagulant and Thrombolytic activities. Regular consumption of L. speciosa leaves may prevent or treat thrombotic Diseases.

Leonurus japonicus Sweet extract

Simultaneous optimization of ultrasonic-assisted extraction of antioxidant and antiCoagulation activities of compounds from Leonurus japonicus Houtt. by response surface methodology

Leonurus japonicus Houtt. is a herbaceous annual of the Lamiaceae family with pantropical distribution; it is called motherwort in China. It plays important roles in anticoagulation and antioxidation. This study aimed to explore the optimization of ultrasound-assisted extraction of multiple compounds from motherwort as well as their antioxidant and antiCoagulation activities using response surface methodology. Box–Behnken design was employed to optimize three significant influences, namely extraction time, extraction temperature and ethanol concentration. The optimum extraction parameters acquired based on a combination of the yield of the target compounds and their antioxidant and anti coagulation activities were an extraction time of 38.2 min, an extraction temperature of 30.0 °C, an ethanol concentration of 48.9% (v/v), a solid–liquid ratio of 30.0 mL g⁻¹ and an ultrasonic power of 500.0 W.

Under the optimal conditions, the maximal yield of the antiCoagulation and antioxidant compounds of motherwort was 0.994%; the thrombin time was 19.872 s; prothrombin time was 8.270 s; the activated partial thromboplastin time was 15.535 s; the fibrinogen was 1.420 g L⁻¹; and the 1,1-diphenyl-2-picrylhydrazyl free radical scavenging activity was 1.503 mg mL⁻¹. The optimized conditions model showed a good correlation between the predicted and experimental values. According to the results of our study, the optimization extraction significantly enabled study of the antiCoagulation and antioxidant activities of compounds in motherwort; this may contribute to future research on the pharmacological activities of motherwort.

Leonurine Ameliorates Oxidative Stress and Insufficient Angiogenesis by Regulating the PI3K/Akt-eNOS Signaling Pathway in H2O2-Induced HUVECs

Thrombus is considered to be the pathological source of morbidity and mortality of Cardiovascular Disease and thrombotic complications, while oxidative stress is regarded as an important factor in vascular endothelial injury and thrombus formation. Therefore, antioxidative stress and maintaining the normal function of vascular endothelial cells are greatly significant in regulating vascular tension and maintaining a nonthrombotic environment. Leonurine (LEO) is a unique alkaloid isolated from Leonurus japonicus Houtt (a traditional Chinese medicine (TCM)), which has shown a good effect on promoting blood circulation and removing blood stasis.

In this study, we explored the protective effect and action mechanism of LEO on human umbilical vein endothelial cells (HUVECs) after damage by hydrogen peroxide (H2O2). The protective effects of LEO on H2O2-induced HUVECs were determined by measuring the cell viability, cell migration, tube formation, and oxidative biomarkers. The underlying mechanism of antioxidation of LEO was investigated by RT-qPCR and western blotting.

Our results showed that LEO treatment promoted cell viability; remarkably downregulated the intracellular generation of reactive oxygen species (ROS), malondialdehyde (MDA) production, and lactate dehydrogenase (LDH); and upregulated the nitric oxide (NO) and superoxide dismutase (SOD) activity in H2O2-induced HUVECs. At the same time, LEO treatment significantly promoted the phosphorylation level of angiogenic protein PI3K, Akt, and eNOS and the expression level of survival factor Bcl2 and decreased the expression level of death factor Bax and caspase3. In conclusion, our findings suggested that LEO can ameliorate the oxidative stress damage and insufficient angiogenesis of HUVECs induced by H2O2 through activating the PI3K/Akt-eNOS signaling pathway.

Licorice root extract

Glycyrrhizin Prevents Hemorrhagic Transformation and Improves Neurological Outcome in Ischemic Diseases Stroke with Delayed thrombolysis Through Targeting Peroxynitrite-Mediated HMGB1 Signaling

Peroxynitrite (ONOO⁻) and high mobility group box 1 protein (HMGB1) are important cytotoxic factors contributing to cerebral ischemia -reperfusion injury. However, the roles of ONOO⁻ in mediating HMGB1 expression and its impacts on hemorrhagic transformation (HT) in Ischemic Diseases brain injury with delayed t-PA treatment remain unclear. In the present study, we tested the hypothesis that ONOO⁻ could directly mediate the activation and release of HMGB1 in Ischemic Diseases brains with delayed t-PA treatment.

With clinical studies, we found that plasma nitrotyrosine (NT, a surrogate marker of ONOO⁻) was positively correlated with HMGB1 level in acute Ischemic Diseases Stroke patients. Hemorrhagic transformation and t-PA-treated Ischemic Diseases Stroke patients had increased levels of nitrotyrosine and HMGB1 in plasma. In animal experiments, we found that FeTmPyP, a representative ONOO⁻ decomposition catalyst (PDC), significantly reduced the expression of HMGB1 and its receptor TLR2, and inhibited MMP-9 activation, preserved collagen IV and tight junction claudin-5 in Ischemic Diseases rat brains with delayed t-PA treatment. ONOO⁻ donor SIN-1 directly induced expression of HMGB1 and its receptor TLR2 in naive rat brains in vivo and induced HMGB1 in brain microvascular endothelial b.End3 cells in vitro.

Those results suggest that ONOO⁻ could activate HMGB1/TLR2/MMP-9 signaling. We then addressed whether glycyrrhizin, a natural HMGB1 inhibitor, could inhibit ONOO⁻ production and the antioxidant properties of glycyrrhizin contribute to the inhibition of HMGB1 and the neuroprotective effects on attenuating hemorrhagic transformation in Ischemic Diseases Stroke with delayed t-PA treatment. Glycyrrhizin treatment downregulated the expressions of NADPH oxidase p47 phox and p67 phox and iNOS, inhibited superoxide and ONOO⁻ production, reduced the expression of HMGB1, TLR2, MMP-9, preserved type IV collagen and claudin-5 in Ischemic Diseases brains.

Furthermore, glycyrrhizin significantly decreased the mortality rate, attenuated hemorrhagic transformation, brain swelling, blood-brain barrier damage, neuronal apoptosis, and improved neurological outcomes in the Ischemic Diseases Stroke rat model with delayed t-PA treatment. In conclusion, peroxynitrite-mediated HMGB1/TLR2 signaling contributes to hemorrhagic transformation, and glycyrrhizin could be a potential adjuvant therapy to attenuate hemorrhagic transformation, possibly through inhibiting the ONOO⁻/HMGB1/TLR2 signaling cascades.

inhibition of human thrombin by the constituents of licorice: inhibition kinetics and mechanistic insights through in vitro and in silico studies

thrombin inhibition therapy is a practical strategy to reduce thrombotic and Cardiovascular risks via blocking the formation of blood Clots. This study aimed to identify naturally occurring thrombin inhibitors from licorice (one of the most popular edible herbs), as well as to investigate their inhibitory mechanisms. Among all tested licorice constituents, licochalcone A was found as the most efficacious agent against human thrombin (IC50 = 7.96 μM). Inhibition kinetic analyses demonstrated that licochalcone A was a mixed inhibitor against thrombin -mediated Z-Gly-Gly-Arg-AMC acetate hydrolysis, with a Ki value of 12.23 μM.

Furthermore, mass spectrometry-based chemoproteomic assays and molecular docking simulations revealed that licochalcone A could bind to human thrombin at both exosite I and the catalytic site. In summary, our findings demonstrated that the chalcones isolated from licorice were a new class of direct thrombin inhibitors, also suggesting that licochalcone A was a promising lead compound for developing novel anti-thrombotic agents.

Nutritional and therapeutic potential of Glycyrrhiza glabra L. roots

This study was aimed to estimate the antioxidant , anti-diabetic, anti-amnesic, anti-biofilm, Thrombolytic, hemolytic, and cytotoxic effects of G. glabra root extracts for nutritive and medicinal purpose. antioxidant, alpha amylase, antiglycation, anti-amnesic, cytotoxic, Thrombolytic and antibiofilm assays were performed.

In the current research, G. glabra root extracts showed significant antioxidant (80.96 mg GAE/100 g TPC, 38.96 mg CE/100 g TFC, 46% DPPH scavenging), antidiabetic (75% antiglycation, 38.7% α-amylase inhibition ), anti-amnesic (5.5%), hemolytic (19.46%), Thrombolytic (35.4%) and antibiofilm (72% P. multocida, 51.57% S. aureus inhibition ) activities. G. glabra root extracts have remarkable activity to fight against oxidative stress, hyperglycemia, blood Coagulation, bacterial growth, DNA damage and memory enhancing ability that ensure its uses in food and pharmaceutical industries.

Possible Pharmacological Basis for antithrombotic Glycyrrhiza Glabra in Sprague Dawley Rats

It has been previously identified that 3-aryl comarin derivative, GU-7 isolated from Liquorice posses Antiplatelet activity. It inhibits platelet aggregation by increasing intraplatelet cyclic AMP concentration. Here we report the in-vivo effects of extract of Glycyrrhiza glabra and also the combined effect with Vitamin K and Heparin.

Extract of Glycyrrhiza glabra increased the bleeding time when given in the doses of 180 mg/kg and 360 mg/kg. blood loss was evaluated 60 minute later as a function of absorbance at 540 nm due to hemoglobin content in water solution. Altogether data indicates that Glycyrrhiza glabra is an effective anti thrombotic agent in vivo, which may account for its known pharmacological properties.

Ligusticum chuam.iong Hort. root extract

Protective effects of Tao-Hong-Si-Wu Decoction on Memory Impairment and Hippocampal Damage in Animal Model of vascular Dementia

Tao-Hong-Si-Wu decoction (TSD) as a traditional chinese medicine (TCM) has been developed to treat thrombotic Diseases for hundreds of years, and vascular dementia (VD) is a cognitive dysfunction syndrome caused by cerebral Embolism . In this study, the protective effect of TSD on memory impairment and brain damage in rat model of VD induced by middle cerebral Artery occlusion (MCAO) was investigated. The study showed that rats in MCAO treatment with TSD for 14 days significantly improved behavioral function, increased densities of neuron, and induced angiogenesis in the brain compared with model rats. TSD also adjusted the neurotransmitter levels, reduced the content of endothelin-1 (ET-1), and induced the activities of vascular endothelial growth factor (VEGF) in hippocampus.

Moreover, the immunohistochemical staining and western blotting results also revealed that TSD decreased apoptosis via upregulated B-cell lymphoma-2 (Bcl-2)/Bcl-2 associated X protein (Bax) ratio. These results demonstrated TSD possesses neuroprotective and antidementia properties by preventing the loss of neural cells, adjusting brain neurotransmitter, promoting cerebral blood circulation, and decreasing apoptosis. These results suggested that TSD might be developed as an effective drug for the prevention of VD.

Establishment of the pharmacological basis of the therapeutic effects of Ligusticum chuanxiong, a traditional Chinese herb for Cardiovascular Diseases

igusticum chuanxiong is a popular Chinese herb for Cardiovascular Diseases in China. However, its use is limited by lack of a scientific foundation. The present study aims to establish the pharmacological basis of the therapeutic effects of Ligusticum chuanxiong. Among 17 major constituents identified in Ligusticum chuanxiong. 6 were orally absorbable as predicted using a Caco-2 colonic cell model. Subsequent screening on vasorelaxation. the most widely examined effect of the herb, showed that absorbable constituents ligustilide. senkyunolide A and butylideneph thalide had relaxing effects comparable to the parent herb. The same three constituents also possessed similar anti-platelet aggregation and anti-thrombotic profiles to the parent herb. Ligusticum chuanxiong was found to have vasorelaxing, platelet -inhibitory and anti-thrombotic actions.

These effects were most likely due to the combined contribution of the three major absorbable constituents ligustilide. senkyunolide A and butylidenephthalide. Key words: Ligusticum chuanxiong, vasorelaxation, platelet, Thrombosis

Lonicerae Japonicae Flos extract

Updated pharmacological effects of Lonicerae japonicae flos, with a focus on its potential efficacy on coronavirus Disease –2019 (Covid -19 )

Lonicerae japonicae flos (LJF), known as Jin Yin Hua in Chinese, is one of the most commonly used traditional Chinese herbs and nutraceuticals. Nowadays, LJF is broadly applied in an array of afflictions, such as fever, sore throat, flu infection, cough, and arthritis, with the action mechanism to be elucidated.

Here, we strove to summarize the main phytochemical components of LJF and review its updated pharmacological effects , including inhibition of inflammation, pyrexia, viruses, and bacteria, immunoregulation, and protection of the liver, nervous system, and Heart, with a focus on the potential efficacy of LJF on coronavirus Disease–2019 based on network pharmacology so as to fully underpin the utilization of LJF as a medicinal herb and a favorable nutraceutical in daily life.

N-Acetylcysteine

Potent Thrombolytic Effect of N-Acetylcysteine on Arterial Thrombi

Background: Platelet cross-linking during Arterial Thrombosis involves von Willebrand Factor (VWF) multimers. Therefore, proteolysis of VWF appears promising to disaggregate platelet -rich thrombi and restore vessel patency in acute thrombotic disorders such as Ischemic Diseases Stroke , acute Coronary syndrome, or acute limb ischemia . N-Acetylcysteine (NAC, a clinically approved mucolytic drug) can reduce intrachain disulfide bonds in large polymeric proteins. In the present study, we postulated that NAC might cleave the VWF multimers inside occlusive thrombi, thereby leading to their dissolution and Arterial recanalization.

Methods: Experimental models of thrombotic Stroke induced by either intra-Arterial thrombin injection or ferric chloride application followed by measurement of cerebral blood flow using a combination of laser Doppler flow metry and MRI were performed to uncover the effects of NAC on Arterial thrombi. To investigate the effect of NAC on larger vessels, we also performed ferric chloride–induced carotid Artery Thrombosis . In vitro experiments were performed to study the molecular bases of NAC Thrombolytic effect, including platelet aggregometry, platelet -rich thrombi lysis assays, thromboelastography (ROTEM), and high-shear VWF string formation using microfluidic devices. We also investigated the putative prohemorrhagic effect of NAC in a mouse model of intracranial hemorrhage induced by in situ collagenase type VII injection.

Results: We demonstrated that intraVenous NAC administration promotes lysis of Arterial thrombi that are resistant to conventional approaches such as recombinant tissue-type plasminogen activator, direct thrombin inhibitors, and antiplatelet treatments. Through in vitro and in vivo experiments, we provide evidence that the molecular target underlying the Thrombolytic effects of NAC is principally the VWF that cross-link platelet s in Arterial thrombi. Coadministration of NAC and a nonpeptidic GpIIb/IIIa inhibitor further improved its Thrombolytic efficacy, essentially by accelerating thrombus dissolution and preventing reThrombosis . Thus, in a new large-vessel Thromboembolic Stroke model in mice, this cotreatment significantly improved Ischemic Diseases lesion size and neurological outcome. It is important to note that NAC did not worsen hemorrhagic Stroke outcome, suggesting that it exerts Thrombolytic effects without significantly impairing normal hemostasis.

Conclusions: We provide evidence that NAC is an effective and safe alternative to currently available antithrombotic agent s to restore vessel patency after Arterial occlusion.

Treatment of refractory thrombotic thrombocytopenic purpura with N-acetylcysteine: a case report

Background: Thrombotic thrombocytopenic purpura (TTP) is a life-threatening Disease resulting in systemic microvascular Thrombosis . The Disease is caused by excessive platelet (PLT) adhesion to ultra-large (UL) von Willebrand factor (VWF) multimers inadequately cleaved by the processing enzyme ADAMTS-13. While many cases respond to plasma exchange performed with or without concurrent corticosteroids, treatment of the 10% to 20% of patients with refractory Disease is difficult. Experimental studies demonstrating that N-acetylcysteine (NAC) inhibits PLT binding to endothelial cell–secreted and anchored UL VWF multimers suggest that NAC may be useful in the treatment of TTP.

Case Report: A 44-year-old woman presented with malaise, confusion, chest and abdominal pain, and transient visual loss. Laboratory results and peripheral blood smear were consistent with TTP. The patient was begun on plasma exchange and corticosteroid treatment, but after 10 days the PLT count was still less than 10.0 × 109/L and she developed a fever. Rituximab was initiated, but the patient’s condition worsened and she became comatose. Antibiotics were initiated, but cultures remained sterile. After 3 days of coma and further clinical deterioration, treatment with NAC was begun. The patient received a loading dose of 150 mg/kg NAC intraVenous ly (IV) over 1 hour. Within 18 hours the patient awakened abruptly and began communicating with medical personnel. Plasma exchange, corticosteroids, rituximab, and NAC infusion (150 mg/kg IV over 17 hr daily × 10 days) were continued and by Day 17 the PLT count was more than 50 × 109/L. The patient fully recovered and was discharged on Day 31.

Conclusion: This is the first complete report of a TTP patient treated with NAC. NAC was a safe and effective supplementary treatment for refractory TTP in this patient.

N-acetylcysteine inhibits Thrombosis in a murine model of myeloproliferative neoplasm

Thrombosis is a major cause of mortality in patients with myeloproliferative neoplasms (MPNs), though there is currently little to offer patients with MPN beyond aspirin and cytoreductive therapies such as hydroxyurea for primary prevention. Thrombogenesis in MPN involves multiple cellular mechanisms, including platelet activation and neutrophil-extracellular trap formation; therefore, an antithrombotic agent that targets one or more of these processes would be of therapeutic benefit in MPN.

Here, we treated the JAK2V617F knockin mouse model of polycythemia vera with N-acetylcysteine (NAC), a sulfhydryl-containing compound with broad effects on glutathione replenishment, free radical scavenging, and reducing disulfide bonds, to investigate its antithrombotic effects in the context of MPN. Strikingly, NAC treatment extended the lifespan of JAK2V617F mice without impacting blood counts or splenomegaly. Using an acute Pulmonary Thrombosis model in vivo, we found that NAC reduced thrombus formation to a similar extent as the irreversible platelet inhibitor aspirin. In vitro analysis of platelet activation revealed that NAC reduced thrombin-induced platelet-leukocyte aggregate formation in JAK2V617F mice.

Furthermore, NAC reduced neutrophil extracellular trap formation in primary human neutrophils from patients with MPN as well as healthy controls. These results provide evidence that N-acetylcysteine inhibits Thrombosis in JAK2V617F mice and provide a pre-clinical rationale for investigating NAC as a therapeutic to reduce thrombotic risk in MPN.

Natto

Thrombolytic Effect of Nattokinase on a Chemically Induced Thrombosis Model in Rat

Nattokinase is a new fibrinolytic enzyme which cleaves directly cross-linked fibrin in vitro. In this study, we investigated the Thrombolytic effect of nattokinase on a thrombus in the common carotid Artery of rat in which the endothelial cells of the vessel wall were injured by acetic acid. When a section of occluded vessel was stained for CD61 antigen by immunofluorescence utilizing a monoclonal antibody, the antigen was localized around the surface of the occluded blood vessels.

This result suggests that the occlusive Thrombosis was caused by platelet aggregation . In addition, thrombolysis with urokinase (UK ; 50000IU/kg, i.v.) or tissue plasminogen activator (tPA ; 13300IU/kg, i.v.) in our model was observed to restore the blood flow over a 60 min monitoring period. The results indicate that our chemically induced model is useful for screening and evaluating a Thrombolytic agent.

We evaluated the Thrombolytic activity of nattokinase using this model and compared it with fibrino (geno) lytic enzyme, plasmin or elastase. On a molar basis, the recovery of the Arterial blood flow with nattokinase, plasmin and elastase were 62.0±5.3%, 15.8±0.7% and 0%, respectively. The results indicate that the Thrombolytic activity of nattokinase is stronger than that of plasmin or elastase in vivo.

Natto and Its Active Ingredient Nattokinase: A Potent and Safe Thrombolytic agent

Nattokinase: An Oral antithrombotic agent for the Prevention of Cardiovascular Disease

Natto, a fermented soybean product, has been consumed as a traditional food in Japan for thousands of years. Nattokinase (NK), a potent blood–Clot dissolving protein used for the treatment of Cardiovascular Diseases, is produced by the bacterium Bacillus subtilis during the fermentation of soybeans to produce Natto. NK has been extensively studied in Japan, Korea, and China. Recently, the fibrinolytic (anti-clotting) capacity of NK has been recognized by Western medicine. The National Science Foundation in the United States has investigated and evaluated the safety of NK. NK is currently undergoing a clinical trial study (Phase II) in the USA for atherothrombotic prevention.

Multiple NK genes have been cloned, characterized, and produced in various expression system studies. Recombinant technology represents a promising approach for the production of NK with high purity for its use in antithrombotic applications. This review covers the history, benefit, safety, and production of NK. Opportunities for utilizing plant systems for the large-scale production of NK, or for the production of edible plants that can be used to provide oral delivery of NK without extraction and purification are also discussed.

Thrombolytic effects in vivo of Nattokinase in a Carrageenan-Induced Rat Model of Thrombosis

Background/Aims: Nattokinase is a serine protease produced by Bacillus subtilis during the fermentation of the soybean product natto. The fibrinolytic activity and Thrombolytic effects of nattokinase have been observed in vitro, but the effect in vivo has still to be researched. The objective of this study was to demonstrate the activity of nattokinase in vivo.

Methods: To establish a rat model of Thrombosis, κ-carrageenan was injected subcutaneously into the toes of Sprague-Dawley (SD) rats. Histological examination confirmed Thrombosis. The rats were then treated with varying doses of nattokinase and the resulting thrombolysis was histologically assessed. ELISA was used to determine the levels of the fibrin/fibrinogen degradation products (FDPs) and D-dimer, which are sensitive indices of fibrinolytic activity. Vermis kinase, a known Thrombolytic agent, was used as a positive control.

Results: Biopsy results revealed partial thrombolysis in the tail vessels of the rats treated with nattokinase or vermis kinase. FDP and D-dimer levels were higher in rats treated with high-dose nattokinase than in those treated with saline. No difference in FDP or D-dimer levels was observed between rats treated with high-dose nattokinase and those treated with vermis kinase.

Conclusions: Both the histological and physiological evidence from this study indicate that nattokinase exerts Thrombolytic effects in vivo.

In vitro and in silico studies on fibrinolytic activity of nattokinase: A Clot buster from Bacillus sp.

Background: Nattokinase (NK) is a serine protease enzyme of the subtilisin family. It exhibits a strong fibrinolytic activity. The fibrinolytic enzymes from Bacillus sp. have attracted interest as Thrombolytic agent s because of their efficiency in the fibrinolytic process including plasmin activation .

Methods: In the present study, VIT garden soil was collected and subjected to isolation process in order to screen for the NK production. Screening for NK enzyme was performed by radial caseinolytic assay. The production of NK enzyme was done in two different production medium for comparative studies. The NK enzyme was purified by gel permeation chromatography. The activity of the purified NK was checked by Clotlysis and casein digestion assay. To investigate the structural basis of NK and fibrinogen interaction and also to identify the best binding mode, molecular dynamics and docking studies were performed.

Results: Based on the morphological and biochemical characterization, the isolate was identified as Bacillus sp. The overall purification fold of NK was about 3 with the specific activity of 664U/mg and 9.9% yield. Homogeneity of the purified enzyme was analyzed and confirmed by the single band obtained in SDS-PAGE. Molecular weight of the purified protease was estimated as 25 kDa. Purified NK enzyme exhibited 97% of effective Clotlysis activity. The NK was docked in to the knob region of the fibrinogen at its binding site using Dock server. A total of 26 residues of fibrinogen and 29 residues of NK constitute the interface region. However, 9 residues of fibrinogen (THR238, MET264, LYS266, ARG275, THR277, ALA279, ASN308, MET310, and LYS321) and 8 residues of NK (GLY61, SER63, THR99, PHE189, LEU209, TYR217, ASN218, and MET222) are involved in intact binding.

Conclusions: A significant amount of NK enzyme was obtained from Bacillus sp. The docking analysis revealed that the NK and fibrinogen adopt an extended binding pattern and interacts with the crucial residues to exhibit their activity.

A Newly Derived Protein From Bacillus subtilis natto With Both antithrombotic and fibrinolytic effects

Natto, steamed soybeans fermented by Bacillus subtilis natto, is a traditional Japanese food. We derived a purified protein layer, called NKCP as a trade mark, from B. subtilis natto fermentation. In the present study, we examined the fibrinolytic and antithrombotic effects of NKCP and identified its active component to clarify the fibrinolytic effect of NKCP observed in preliminary clinical trials previously. The active component of NKCP was identified as a 34-kilodalton protein designated bacillopeptidase F. NKCP showed direct degradation of artificial blood Clot in saline. The protease activity, accounting for the fibrinolytic effect of NKCP, was examined with a chromogenic substrate for plasmin. Dose-dependent prolongations of both prothrombin time and active partial thromboplastin time were observed in rats with intraduodenum administration of NKCP.

Our in vitro and in vivo studies suggest that NKCP has both a fibrinolytic effect and an antithrombotic effect similar to heparin. Because NKCP is derived from food and has safety data demonstrated by previous animal experiments and preliminary clinical trials, NKCP is considered as safe for clinical use.

Nelumbinis Stamen (Lotus)

Plumula Nelumbinis: A review of traditional uses, phytochemistry, pharmacology, pharmacokinetics and safety

Results: Plumula Nelumbinis is used to treat hypertension , arrhythmia, severe aplastic anemia, insomnia, encephalopathy and gynecological Disease in traditional Chinese medicine and clinical studies. More than 130 chemicals have been isolated and identified from Plumula Nelumbinis, including alkaloids, flavonoids, polysaccharides and volatile oil. In addition, pharmacological effects , such as protective effects against Cardiovascular Diseases , neurological Diseases , lung and kidney injury, anti-inflammatory and anticancer activities, were also evaluated by in vitro and in vivo studies. Moreover, the potential signaling pathways regulated by Plumula Nelumbinis in Cardiovascular and neurological Diseases and perspectives on Plumula Nelumbinis research were discussed.

Systematic chemical analysis of flavonoids in the Nelumbinis stamen

The stamen of lotus, known as Nelumbinis stamen, has been used as the folk medicine and functional food for a long time, which showed good activities of anti-ulcer, AntiThrombosis , analgesic, anti-diarrhea, strengthen uterine contraction. The bioactivities of Nelumbinis stamen were attributed to the existence of flavonoids, its characteristic chemical constituents. A reliable method for comprehensive chemical analysis of flavonoids in Nelumbinis stamen by HPLC–DAD–MS was developed for the first time. The extraction protocol of flavonoids from Nelumbinis stamen was optimized by an orthogonal design.

The chromatographic conditions were optimized, which exhibited similar level than that of the UHPLC platform allowing target compound identification in a shorter time with little solvent consumption. Moreover, similarity analysis, hierarchical clustering analysis and principal components analysis were successfully applied to demonstrate the variability of these Nelumbinis stamen samples.

Nigella sativa

The Thrombolytic and Cytotoxic effects of Nigella sativa (L.) Seeds: The Prophetic Medicine

The Water-Soluble Extract (WSE) is a crude bioactive phytoconstituent of Nigella sativa (L.) seeds discovered recently. The current findings report about the Thrombolytic and cytotoxic effects of WSE using human blood Clotlysis and brine shrimp lethality (BSL) bioassay. The Thrombolytic effect of WSE (1,666.67 µg/mL) was determined via the Clot and lysate weight measurements compared to streptokinase (STK) of 30,000 IU/mL and normal saline (NS) while the cytotoxicity of WSE (44.14-2,000 µg/mL) against vincristine sulfate (VCS;3.125-100 µg/mL). WSE has shown extremely statistically significant (p<0.0001) Clotlysis (90.00%) compared to NS (3.76%) whilst it was also significantly different (p<0.0063) to STK (72.41%) exhibiting LC50 of 1,795.90 µg/mL vs. VCS (39.25 µg/mL) in a dose-dependent manner.

The current results suggested WSE has a potent Thrombolytic effect with mild dose-dependent cytotoxicity towards brine shrimp nauplii (Artemia salina). It also suggested WSE might have enzymatic roles on thrombin, fibrin, and plasmin of blood. This pharmacological action of WSE is might be due to its antioxidant property, short-chain fatty acids and/or amino acids. Further studies are highly recommended on the enzymatic role(s) and bioactive phytoconstituents of WSE.

In vitro induction of endothelial cell fibrinolytic alterations by Nigella sativa

The effect of Nigella sativa (NS) L. oil (blackseed oil) on the fibrinolytic system of the human umbilical vein (HUV) and human uterine Arterial (HUA) endothelial cells (ECs) in culture was studied. Both of them showed a concentration-dependent increase in tissue-type plasminogen activator (t-PA). A maximum effect was achieved with 50 μg oil/ml conditioned medium (CM) (1.3±0.15 ng/104 cells/24 h vs. control 0.7±0.06 ng/104 cells/24 h, and 0.38±0.04 ng/104 cells/24 h vs. control 0.24±0.02 ng/104 cells/24 h, for HUVEC and HUA-EC, respectively).

At 100 μg/ml, there was a significant change in the amount of t-PA antigen produced by either HUVEC or HUA-EC (1.0±0.1 ng/104 cells/24 h or 0.28±0.02 ng/104 cells/24 h) as compared to control CM from cells grown under control conditions, but still less than that recorded at 50 μg oil/ml. Plasminogen activator inhibitor-type 1 increased the CM significantly and concentration-dependently in both cells. For HUVEC, the maximum effect was achieved at a concentration of 100 μg/ml (257.7±8.0 ng/104 cells/24 h vs. control 72.7±3.8 ng/104 cells/24 h). HUA-EC showed the maximum effect at a concentration of 100 μg/ml (171.6±4.4 ng/104 cells/24 h vs. control 53.8±3.7 ng/104 cells/24 h). This study suggests a role for NS oil in modulating the balance of fibrinolysis/thrombus formation by modulating the fibrinolytic potential of endothelial cells.

Evaluation of Thrombolytic properties of Nigella sativa, Capsicum frutescens and Brassica oleracea

Thrombolytic drugs are widely used for the management of cerebral Venous sinus Thrombosis patients. Several in vitro models have been developed to study Clotlysis activity of Thrombolytic drugs, but all of these have certain limitations. The present study was carried out to investigate the Thrombolytic activity, synergistic activity on thrombolysis in the aqueous extract of fruits of Nigella sativa, Capsicum frutescens, Brassica oleracea. An in vitro Thrombolytic method was used to investigate the Thrombolytic activity of plant extracts in blood sample from healthy human volunteers, along with streptokinase as a positive control and water as a negative control. Same method was also applied to assess the synergistic activity of plant extracts with streptokinase. Aqueous extract of Nigella sativa, Capsicum frutescens, Brassica oleracea showed 40.65%, 36.93%, 57.03% Thrombolytic activity and also showed 60.46%, 80.00%, 72.50% synergistic activity with streptokinase respectively.

Among the plant extracts studied Capsicum frutescens showed significant % of synergistic activity (80%) with compared to other plant extracts. The present study demonstrates that Capsicum frutescens extract showed synergistic activity with streptokinase on thrombolysis. Isolation and purification of the compound responsible for synergistic activity would be one of the best sources of herbal drugs for atherothrombotic Diseases.

Paeonia suffruticosa Andr. peel extract

antithrombotic effects of Paeoniflorin from Paeonia suffruticosa by Selective inhibition on Shear Stress-Induced platelet aggregation

antiplatelet agent s are important in the pharmacotherapeutic regime for many Cardiovascular Diseases , including thrombotic disorders . However, bleeding, the most serious adverse effect associated with current antiplatelet therapy, has led to many efforts to discover novel anti-platelet drugs without bleeding issues. Of note, shear stress-induced platelet aggregation (SIPA) is a promising target to overcome bleeding since SIPA happens only in pathological conditions. Accordingly, this study was carried out to discover antiplatelet agent s selectively targeting SIPA. By screening various herbal extracts, Paeonia suffruticosa and its major bioactive constituent, paeoniflorin, were identified to have significant inhibitory effects against shear-induced aggregation in human platelets.

The effects of paeoniflorin on intraplatelet calcium levels, platelet degranulation, and integrin activation in high shear stress conditions were evaluated by a range of in vitro experiments using human platelets. The inhibitory effect of paeoniflorin was determined to be highly selective against SIPA, through modulating von Willebrand Factor (vWF)-platelet glycoprotein Ib (GP Ib) interaction. The effects of paeoniflorin on platelet functions under high shear stress were confirmed in the ex vivo SIPA models in rats, showing the good accordance with the anti-SIPA effects on human platelets. Treatment with paeoniflorin significantly prevented Arterial Thrombosis in vivo from the dose of 10 mg/kg without prolonging bleeding time or blood Clot ting time in rats. Collectively, our results demonstrated that paeoniflorin can be a novel anti-platelet agent selectively targeting SIPA with an improved safety profile.

platelet anti-aggregatory and blood anti-coagulant effects of compounds isolated from Paeonia lactiflora and Paeonia suffruticosa

The roots of two Paeoniaceae family members have long been used as traditional medicines in Korea, China, and Japan. Dry roots of Paeonia lactiflora and dry root bark of P. suffruticosa are used under the traditional names of Paeoniae Radix and Moutan Cortex, respectively. Both Paeoniae Radix and Moutan Cortex have been used as remedies for Cardiovascular Diseases , for improving blood circulation, or for other uses. It was postulated that both plants may contain common active constituents that contribute to inhibiting blood Coagulation and/or platelet aggregation. Eighteen compounds, which have been reported to be present in both plant medicines, were evaluated for their effects on platelet aggregation and blood Coagulation.

Paeonol (5), paeoniflorin (9), benzoylpaeoniflorin (11), and benzoyloxypaeoniflorin (12) were found to be the major common active constituents and they would collectively contribute to improving blood circulation through their inhibitory effects on both platelet aggregation and blood Coagulation. In addition, methylgallate (4), (+)-catechin (7), paeoniflorigenone (8), galloylpaeoniflorin (13), and daucosterol (16) may also take part in improving blood circulation by inhibiting ether platelet aggregation and/or blood Coagulation.

Paeonia veitchii Lynch root extract

Naoxintong capsule limits Myocardial infarct expansion by inhibiting platelet activation through the ERK5 pathway

Purpose: This study investigated whether NXT could affect platelet function and activation under Ischemic Diseases pathological conditions.

Materials and methods: Wistar rats were divided into six groups, sham, saline, NXT (250, 500, 1000 mg/kg), and aspirin group (10 mg/kg). For the pre-treatment assays, MI model was established after pre-administration of saline, NXT-L, NXT-M, NXT-H, and aspirin respectively for 14 days, and after surgery, there were no continuous treatments. For the post-treatment assay, rats were orally administered for 3 days after MI. FeCl₃-induced Thrombosis model was applied to determine the thrombus wet weight. Bleeding time was used to assess the ability of the platelet s to develop a hemostatic plug.

Results: NXT decreased infarct size, decreased LDH, CK, and CK-MB values, and improved Cardiac function. NXT inhibited platelet s activation through reducing CD62P-positive platelet s and inhibited infarct expansion by decreasing the number of CD45-positive cells and the amount of MMP9 secreted into the Heart tissue. Mechanistically, NXT inhibited platelet s activation through decreasing ROS levels, decreasing ERK5 phosphorylation, and increasing RAC1 phosphorylation in MI rats. Pre-treatment with NXT decreased thrombus formation and had normal bleeding times.

Cellular and Molecular Mechanisms of Antithrombogenic Plants: A Narrative Review

Heart attack, Stroke, and Deep vein Thrombosis are among the conditions that alter blood Coagulation and are modulated by antithrombogenic drugs. Natural products are an important source of antithrombogenic agent s and have been considered remarkable alternatives with greater efficacy and usually with fewer side effects. However, the efficacy and toxicity of many of these plants that are used in traditional medicine must be scientifically tested. Despite a large number of published articles that report that plants or plant-derived components may act as antithrombogenic agents, few studies have investigated the mechanism of action of medicinal plants.

This review presents the current knowledge about the major cellular and molecular mechanisms of antithrombogenic plants and their main components. Many well-established mechanisms (e.g., platelet aggregation, Coagulation factors, and thrombolysis ) are related to the antithrombogenic activity of many natural products. However, the central pathways that are responsible for their activity remain unclear.

Further studies are needed to clarify the central role of each of these pathways in the pleiotropic response to these agents.

Panax notoginseng (Burk.)F. H. Chen root extract

Panax notoginseng saponins reduce high-risk factors for Thrombosis through peroxisome proliferator-activated receptor -γ pathway

The classic Virchow theory suggests that blood stasis, hypercoagulability and endothelial dysfunction are three major factors that cause Venous Thrombosis (VT). It is a complicated biological process involved multi-factors. platelet plays a central role and participates in multiple links of this process. Panax notoginseng saponins (PNS), the principal constituents derived from panax notoginseng, has been widely described for its anti-platelet activity. However, its potential mechanism against platelet aggregation has not been clarified. In this present study, we evaluated the anti-platelet effects of PNS on thrombin -induced platelet activation and its possible molecular mechanism of action, and further explored the therapeutic action of PNS on thrombin induced hypercoagulability in rat.

Our results showed that PNS treatment inhibited platelet aggregation induced by thrombin, which was accompanied with over-expression of Peroxisome proliferator-activated receptor γ (PPAR-γ) protein, mRNA and upregulation of phosphatidylinositol 3 kinase (PI3K)/ protein kinase B (Akt)/ endothelial nitric oxide synthase (eNOS) pathway in platelet, and this effect could be reversed by PPAR-γ inhibitor T0070907.

In vivo, PNS significantly reversed thrombin -induced hypercoagulable state in rat which was accompanied by PPAR-γ protein and mRNA upregulation in rat lung. In conclusion, these data suggested that PNS could suppress thrombin-induced platelet aggregation in vitro and effectively improve hypercoagulable state in vivo and PNS-induced activation of PPAR-γ and its downstream PI3K/Akt/eNOS pathway played the central role.

Panax Notoginseng Saponins Combined with Dual Antiplatelet Drugs Potentiates Anti-Thrombotic Effect with Alleviated Gastric Injury in A Carotid Artery Thrombosis Rat Model

Objective

To observe the combination effects of Panax notoginseng saponins (PNS)and dual antiplatelet drugs (DAPT), and to explore the mechanism via cyclooxygenase /prostaglandin pathway.
Methods

Right carotid Artery Thrombosis was induced in Wistar rats by infiltration with 70% FeCl3, and the animals were randomly divided into sham group, model group, DAPT group and PNS + DAPT group, intragastrically treated for 4 weeks. The cerebral pia mater microcirculation was observed in vivo after anesthetizing by anatomical microscope. The wet weight of carotid Artery Thrombosis was measured. Gastric mucosal injury was observed by hematoxylin and eosin staining. platelet aggregation rate was detected with adenosine diphosphate -induced turbidimetry. platelet CD62p expression was detected by flow cytometry. Concentrations of 6-Ketoprostaglandin F1 alpha, prostaglandin E2 in gastric mucosa and thromboxane B2, 6-Ketoprostaglandin F1 alpha, tissue plasminogen activator, plasminogen activator inhibitor, and fibrin fragment D in the plasma were measured by radioimmunoassay.
Results

PNS and DAPT increased the blood flow volume of cerebral pia mater and decreased erythrocyte aggregation and leukocyte adhesion of model rats. Compared to DAPT, PNS and DAPT further reduced the weight of carotid Artery Thrombosis with enhanced inhibition of platelet aggregation , increased tissue plasminogen activator levels and decreased fibrin fragment D levels. PNS and DAPT alleviated gastric injury induced by dual antiplatelet drugs and upregulated the expression of 6-Ketoprostaglandin F1 alpha in the gastric mucosa compared with DAPT.
Conclusions

PNS combined with DAPT increased anti-Thrombosis effects of DAPT and mitigated DAPT-related gastric injury. The underlying mechanisms may be associated with enhanced antiplatelet aggregation and activation of the fibrinolytic system and up-regulation of 6-Ketoprostaglandin F1 alpha expression in gastric mucosa.

Antiplatelet and Anticoagulant effects of Panax notoginseng: Comparison of raw and steamed Panax notoginseng with Panax ginseng and Panax quinquefolium

Ethnopharmacological significance: Panax notoginseng (Burk.) F. H. Chen (Araliacea) is traditionally used for its hemostatic and Cardiovascular effects when raw and as a tonic when steamed.

Aim of the study: This study aims to compare the effects of raw and steamed Panax notoginseng, Panax ginseng C. A. Meyer and Panax quinquefolium Linn. on platelet aggregation and plasma Coagulation.

Materials and methods: Effects on collagen-induced platelet aggregation were investigated using a platelet aggregometer, while the plasma Coagulation times (prothrombin time, activated partial thromboplastin time and thrombin time) were determined using a blood Coagulation analyzer. The data was corroborated with ex vivo platelet aggregation and in vivo rat bleeding time.

Results: Raw and steamed Panax notoginseng significantly inhibit platelet aggregation and plasma Coagulation. Steamed Panax notoginseng has significantly more potent antiplatelet and Anticoagulant effects than the raw extract, and the antiplatelet and Anticoagulant effects increase with increasing steaming durations. Comparing the three common Panax species, Panax notoginseng has higher antiplatelet effect than Panax ginseng and Panax quinquefolium. The in vitro antiplatelet and Anticoagulant effects are positively translated into a prolongation of in vivo rat bleeding time after oral administration of the raw and steamed extracts.

Conclusion: The results indicate that the three common Panax species affect platelet aggregation and plasma Coagulation differently, with steamed Panax notoginseng showing the greatest antiplatelet and Anticoagulant effects . Panax notoginseng may be a good source of lead compounds for novel antiplatelet and Anticoagulant therapeutics.

Panax notoginseng saponin is superior to aspirin in inhibiting platelet adhesion to injured endothelial cells through COX pathway in vitro

Objective: This study was designed to investigate the effect of Panax notoginseng saponin (PNS) on platelet adhesion to injured endothelial cells (ECs) and platelet activation induced by injured ECs, and to explore its underlying mechanisms.

Methods: Human umbilical vein endothelial cells (HUVECs) pretreated with aspirin (ASA,15 μg/mL) or PNS (160 μg/mL), or neither, were exposed to oxidized low-density lipoprotein (ox-LDL,80 mg/L) for 16 h. platelet s were then added and co-cultured with HUVECs for 5 min. platelet adhesion to ECs, platelet CD62p expression, and HUVEC apoptosis were assessed by fluorescence activated cell sorting (FACS)·Supernatant concentration of 6-keto-PGF1α and thromboxane 2 (TXB2) were measured by radioimmunoassay. Cyclooxygenase-1 (COX-1) and COX-2 protein expression were measured by western blotting.

Results: The inhibitory effect of PNS on platelet activation was similar to ASA, but the inhibitory effect of PNS on platelet adhesion to ECs was superior to ASA. PNS modulated COX-2 expression, and increased 6-keto-PGF1α concentration in HUVECs, while down-regulated COX-1 expression and decreased supernatant TXB2 concentration in platelet s. Co-culturing of injured HUVECs with platelet s increased HUVEC apoptosis induced by ox-LDL compared with HUVECs cultured without platelet s; ASA increased HUVEC apoptosis induced by ox-LDL when cultured without platelets, while decreased the apoptosis when co-cultured with platelets.

Conclusions: EC protection by ASA is closely associated with its inhibitory effect on platelet activation . PNS is superior to ASA in protecting ECs and in inhibiting platelet adhesion to injured ECs, and the regulation of COX pathway in both ECs and platelet s might be the underlying mechanisms of PNS.

Panax ginseng

Covid -19 and Panax ginseng: Targeting platelet aggregation , Thrombosis and the Coagulation pathway

Coronavirus Disease 2019 (Covid -19 ) not only targets the respiratory system but also triggers a cytokine storm and a series of complications, such as gastrointestinal problems, acute kidney injury, and Myocardial ischemia . The use of natural products has been utilized to ease the symptoms of Covid -19 , and in some cases, to strengthen the immune system against Covid -19. Natural products are readily available and have been regularly consumed for various health benefits. Covid -19 has been reported to be associated with the risk of thromboEmbolism and Deep vein Thrombosis.

These thrombotic complications often affects mortality and morbidity. Panax ginseng, which has been widely consumed for its various health benefits has also been reported for its therapeutic effects against Cardiovascular Disease, Thrombosis and platelet aggregation . In this review, we propose that P. ginseng can be consumed as a supplementation against the various associated complications of Covid -19, especially against Thrombosis.

We utilized the network pharmacology approach to validate the potential therapeutic properties of P. ginseng against Covid -19 mediated Thrombosis , the Coagulation pathway and platelet aggregation. Additionally, we aimed to investigate the roles of P. ginseng against Covid -19 with the involvement of platelet -leukocyte aggregates in relation to immunity-related responses in Covid -19.

Possibility as role of ginseng and ginsenosides on inhibiting the Heart Disease of Covid -19 : A systematic review

Coronavirus has been spreading rapidly around the world since it broke out in China in 2019. Respiratory Diseases caused by coronavirus infection cause various Diseases ranging from asymptomatic subclinical infections to severe pneumonia and Cardiovascular complications, leading to death. In this regard, natural products are being studied to prevent various Diseases caused by Covid -19.

In current review, we would like to present mechanisms related to the inhibition of Heart Disease in ginseng and ginsenoside against SARS-CoV-2. In many previous studies, ginseng and ginsenoside are known to have antioxidant, blood flow improvement, improvement of vascular and Heart function, blood pressure control, suppression of Myocardial Infarction and Heart failure, and antiarrhythmia. Therefore, ginseng and ginsenoside have a possibility to suppress Cardiovascular complications caused by Covid -19. Many of research provide evidence for ginseng and ginsenoside as treatments for the risk of Cardiovascular complications.

However, in this review, more specific contents on the proposition of the efficacy of ginseng and ginsenoside for Covid -19 should be presented. Therefore, we hope that researches to reduce Cardiovascular complications of ginseng and ginsenoside for Covid -19 should be presented to reduce mortality for Covid -19.

Potential benefits of ginseng against Covid -19 by targeting inflammasomes

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2 ) is the pathogenic virus that causes coronavirus Disease 2019 (Covid -19 ), with major symptoms including hyper-inflammation and cytokine storm, which consequently impairs the respiratory system and multiple organs, or even cause death. SARS-CoV-2 activates inflammasomes and inflammasome-mediated inflammatory signaling pathways, which are key determinants of hyperinflammation and cytokine storm in Covid -19 patients.

Additionally, SARS-CoV-2 inhibits inflammasome activation to evade the host’s antiviral immunity. Therefore, regulating inflammasome initiation has received increasing attention as a preventive measure in Covid -19 patients. Ginseng and its major active constituents, ginsenosides and saponins, improve the immune system and exert anti-inflammatory effects by targeting inflammasome stimulation. Therefore, this review discussed the potential preventive and therapeutic roles of ginseng in Covid -19 based on its regulatory role in inflammasome initiation and the host’s antiviral immunity.