• Journal of Life Science
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• v.20 no.6
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• pp.922-928
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• 2010

This study was carried out to develop safe and novel anticoagulation agents from oriental medicinal herbs. From 33 medicinal herbs, 40 different ethanol extracts were prepared according to place of origin or extraction parts, and anticoagulation activities were evaluated by determination of thrombin time (TT), prothrombin time (PT) and activated partial thromboplastin time (aPTT). The average water content and average extraction ratio for the medicinal herbs were $6.85{\pm}2.26%$ and $5.27{\pm}4.25%$, respectively. Evaluation of TT at various concentrations of the extract led to the selection of Mucuna birdwoodiana, Prunus armeniaca, Cacalia ainsliaeiflora, Cinnamonum aromaticum, and Rhus javanica Linneas potent antithrombosis medicinal herbs. Evaluation of PT and aPTT showed that the extracts of R.javanica Linne, M. birdwoodiana, and P. armeniaca have strong anticoagulation activities. Determination of hemolytic activities of 40 different ethanol extracts against human red blood cells, however, showed that only M. birdwoodiana, C. ainsliaeiflora, C. aromaticum, and R. javanica Linnehas strong anticoagulation activity without hemolytic activity at a concentration of 500 mg/ml. Our results suggest that oriental medicinal herbs, which are under a mass-production system, have potentialas a safe and novel source of anticoagulants, as well being a thrombin-specific and coagulation factor-specific inhibitor.

• Toxicological Research
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• v.20 no.2
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• pp.137-142
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• 2004

We investigated the effects of plant vinegar on platelets and blood coagulation system. Plant vinegar inhibited in vitro platelet aggregation in a concentration dependent manner, when platelets were activated by thrombin and collagen. In addition, plant vinegar showed inhibitory effects on the serotonin secretion induced by thrombin in a concentration dependent manner. However, treatment with plant vinegar to platelets did not induce any cytotoxicity, as determined by the release of lactate dehydrogenase. Plant vinegar did not change the coagulation parameters such as activated partial thromboplastin time (aPTT) and prothrombin time (PT) using rat citrated plasma. In vivo study revealed that, treatment with plant vinegar prolonged the bleeding time from mouse tail. All these results suggest that plant vinegar might improve blood hemostasis mediated via anti platelet activities.

• Journal of the Korean Society of Food Science and Nutrition
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• v.31 no.5
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• pp.917-923
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• 2002

Inhibitory mechanism of the anticoagulant polysaccharide purified from Codium fragile was investigated. The anticoagulant compounds (Cf-30-IV-4-ii, CF-30-IV) prolonged the clotting time at both activated partial thrombo-plastin time (aPTT) and thrombin time (TT). The Inhibition factor assay of instrinsic coagulation pathway in the blood showed that the anticoagulant polysaccharide (CF-30-IV-4-ii) inhibited other factors such as Ⅷ, Ⅸ, Ⅵ and Ⅷ of the coagulation cascade, which did not affect the lupus anticoagulant AB activity. In the thrombin inhibition pattern the CF-30-IV-4-ii did not directly influence the fibrine formation mediated by thrombin but af-fected the anticoagulant activity through the activation of antithrombin III. Base on these result, the anticoaglant polysaccharide (CF-30-IV-4-ii) was considered to inhibit serine pretense involved in the blood coagulation cascade through the enhancing antithrombin III activity. The residual effects of anticoagulant activity and antithrombosis were tested with ICR mice. The anticoagulant polysaccharide (CF-30-W) kept its anticoagulant activitv for 6 hrs with 100% survival at a dose of 150 mg/kg in the antithromboisis test. The anticoagulant effect of CF-30-RF in ex vivo was proportional to the concentration of intravenously injected dose up to 100 mg/kg.

• Korean Journal of Veterinary Research
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• v.35 no.3
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• pp.595-601
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• 1995

Hemodynamic values were assessed in the cows diseased with mastitis. Hemodynamic testes were performed for white blood cell(WBC), red blood cell(RBC), packed cell volume(PCV), hemoglobin concentration, monocyte, eosinophil, neutrophil, lymphocyte, prothrombin time, activated partial thromboplastin time, thrombin time, fibrinogen, platelet concentrations, antithrombin-III and plasminogen activities. Significant alterations were observed in the mean values of most analytes. The numbers of monocytes, eosinophil, and neutrophil, and prothrombin time were increased while the number of lymphocyte, activated partial thromboplastin time, thrombin time, fibrinogen concentration, plasminogen activity and platelet concentration were decreased. The number of RBC, PCV, hemoglobin and antithrombin-III activity were unchanged compared with normal mean values. These data indicated that activation of hemodynamic mechanisms was initiated either directly by the endotoxin-releasing or indirectly by the inflammatory mediators released by response to etiologic agents. We suspected that the changes of hemodynamic values in the cows diseased with mastitis were very similar to those of experimental endotoxin-induced mastitis.

• BMB Reports
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• v.40 no.5
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• pp.832-838
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• 2007

A novel inhibitory protein against blood coagulation factor Va (FVa) was purified from muscle protein of granulated ark (Tegillarca granosa, order Arcoida, marine bivalvia) by consecutive FPLC method using anion exchange and gel permeation chromatography. In the results of ESI-QTOF tandem mass analysis and database research, it was revealed that the purified T. granosa anticoagulant protein (TGAP) has 7.7 kDa of molecular mass and its partial sequence, HTHLQRAPHPNALGYHGK, has a high identity (64%) with serine/threonine kinase derived from Rhodopirellula baltica (order Planctomycetales, marine bacteria). TGAP could potently prolong thrombin time (TT), corresponding to inhibition of thrombin (FIIa) formation. Specific factor inhibitory assay showed that TGAP inhibits FVa among the major components of prothrombinase complex. In vitro assay for direct-binding affinity using surface plasmon resonance (SPR) spectrometer indicated that TGAP could be directly bound with FVa. In addition, the binding affinity of FVa to FII was decreased by addition of TGAP in dose-dependant manner ($IC_{50}$ value = 77.9 nM). These results illustrated that TGAP might interact with a heavy chain of FVa ($FVa_H$) bound to FII in prothrombin complex. The present study elucidated that non-cytotoxic T. granosa anticoagulant protein (TGAP) bound to FVa can prolong blood coagulation time by inhibiting conversion of FII to FIIa in blood coagulation cascade. In addition, TGAP did not significantly (P < 0.05) show fibrinolytic activity and cytotoxicity on venous endothelial cell line (ECV 304).

• Proceedings of the Korean Society of Applied Pharmacology
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• 1998.11a
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• pp.177-177
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• 1998

In the previous report, tetrandrine (TET) and fangchinoline (FAN) showed antithrombotic and antiplatelet aggregation activities. The present study was undertaken to investigate the effects of tetrandrine and fangchinoline on human platelet aggregation, formation of thromboxane B$_2$ and coagulation of platelet poor plasma. TET and FAN inhibited platelet activating factor (PAF) induced human platelet aggregation, but didn't inhibit the specific binding of PAF to its receptor. Meanwhile, TET and FAN also inhibited PAF, thrombin and arachidonic acid induced thromboxane B$_2$ formation in human washed platelets. In addition, neither TET nor FAN showed any anticoagulation activities in the measurement of the activated partial thromboplastin time (APTT), prothrombin time (PT) and thrombin time (TT) using human platelet poor plasma. These results suggest that antithrombotic effects of TET and FAN in mice may be mainly related to the antiplatelet aggregation activities, and the antiplatelet aggregation effects may be related to the intracellular messenger system such as TXA$_2$ formation etc., but not to the binding of PAF to PAF-receptor on the platelet membrane directly.

• Proceedings of the KOSOMBE Conference
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• v.1997 no.11
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• pp.347-349
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• 1997

In this study, we designed a vascular occlusion device based on the principle that slowing blood flow would induce thrombosis. Tungsten, stainless steel and platinum were tested for finding a good embolic metallic coil. The primary coil and the second coil were done with heat treatment or different time. The pitch distance and the shape retention capability of second coil were characterized by SEM. To increase thrombogenicity, we tried to treat different proteins on steel coils: thrombin, gelatin and both gelatin and thrombin. To verify protein materials treated on coils, the surface of coil treated with different proteins were characterized using Laser Raman Spectrophotometer. After observation of embolic coils, the peptides bonds on theirs' surface were found. In order to compare the thrombogenicity of different embolic agents, we measured whole blood clotting time.

• Journal of the Korean Society of Food Science and Nutrition
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• v.29 no.2
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• pp.335-341
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• 2000

In order to search for blood anticoagulant substance from herbal extracts, we extracted with cold water (Fr. I), methanol (Fr. II) and hot water (Fr. III) from 93 commercially available herbs. The anticoagulant activity of herbal extracts was examined through the intrinsic pathway by activated partial thromboplastin time (APTT), the extriansic pathway by prothrombin time (PT) and the common pathway by thrombin time (TT). Onion, garlic, clove, fenugreek, mugwort, thyme and sage had on anticoagulant activity with intrinsic pathway and mint, italian seasoning, rosemary, turmeric, tarragon and wasabie had on anticoagulant activity with common pathway. Nobody had on anticogulant activity with extrinsic pathway. Among commerical herbs tested, clove cultivated in holland was selected because it showed the most potent anticoagulant activity among the samples investigated.

• Microbiology and Biotechnology Letters
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• v.42 no.3
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• pp.302-306
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• 2014

Ehwa nuruk (EN), a traditional Korean alcoholic rice beverage, is manufactured from pulverized wet rice and the needles of pine trees. In this study, the ethanol extract of EN and its subsequent organic solvent fractions were prepared, and their in-vitro anti-thrombosis activity evaluated. In an anti-coagulation assay, only the ethylacetate (EA) fraction of the ethanol extract showed significant extensions of thrombin time, prothrombin time and activated partial thromboplastin time. In a platelet aggregation assay, the water residue of the ethanol extract exhibited aggregation inhibitory activity. Our results suggest that the EA fraction has potential as a new anticoagulation agent and EN could be used as a novel resource for anti-thrombosis agents. This report provides the first evidence of the anti-thrombosis activity of EN.

• BMB Reports
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• v.29 no.6
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• pp.500-506
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• 1996

We have isolated a water-extracted novel regulator for blood coagulation from an earthworm, Lumbricus rubellus. As a folk remedy, the earthworm has been known to facilitate blood circulation. After complete heat inactivation of endogenous proteases in the earthworm, an anticoagulant(s) was purified through ammonium sulfate fractionation and three consecutive gel permeation chromatography of Sephacryl S-300, Sephadex G-75, and G-150 by measuring activated partial thromboplastin time (APTT) The anticoagulant was further purified to 2,800 fold with a C4 reversed-phase HPLC This activity was stable under heat ($100^{\circ}C$ for 30 min) and acidic conditions (0.4 N HCl). The effects of this partially purified anticoagulant on thrombin were observed with various substrates such as N${\alpha}$-benzoyl-DL-arginine-p-nitroanilide (BApNA), H-D-phenylalanyl-L-pipecoyl-L-arginine-p-nitroanilide (S-2238), N${\alpha}$-p-tosyl-L-arginine methyl ester (TAME), and fibrinogen as a natural substrate. Only TAME hydrolysis, due to an esterase activity of the enzyme, was inhibited among the chromogenic substrates. In addition, the anticoagulant not only inhibited the conversion of fibrinogen to fibrin but also prolonged the fibrin clot formation monitored with the in vitro coagulation test. Based on these observations, we suggest the significance of measuring the ability of antithrombotic drugs to inhibit the esterase activity of thrombin. In this report, it was also shown that the earthworm indeed contained a water-extractable, heat- and acid-stable anticoagulant which could be used as a novel antithrombotic agent.