• Korean Journal of Food Science and Technology
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• v.29 no.4
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• pp.817-822
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• 1997

Inhibitory mechanism of the anticoagulant polysaccharide purified from the fruit body of Coriolus versicolor was investigated in this paper. The anticoagulant polysaccharide (CV-40-Va-1) was proposed to have functions of the inhibition of intrinsic pathway in the blood coagulation pathway together with the interuption of a human platelet aggregation induced by von Willebrand factor (vWF). CV-40-Va-I inhibited other factors of the coagulation cascade such as factor VIII, IX, and as well as thrombin. Especially, CV-40-Va-I inhibited the fibrin formation mediated by thromin, however the polysaccharide did not affect the fibrin formation directly but affected the anticoagulant activity through the activation of antithrombin III. The sulfation of the anticoagulant polysaccharide increased the anticoagulant activity, showing that the sulfate concentration of anticoagulant polysaccharide was important factor in the blood coagulation cascade. Low molecular weight subfraction (MW 1,000) obtained by partial hydrolysis of the CV-40-Va-1 generated potent antiplatelet activity, but showed decreased anticoagulant activity.

• 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.

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2000.05a
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• pp.90-91
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• 2000

The physiological systems that control blood fluidity are both complex and elegant. Blood must remain fluid within the vasculature and yet clot quickly when exposed to nonendothelial surfaces at sites of vascular injury. There are two principle mechanisms to control a delicate balance in higher organisms (Davie & Ratnoff, 1964). Present evidence suggests that the intrinsic pathway play an important role in the growth and maintenance of fibrin formation in the coagulation cascade while a second overlapping mechanism, called the extrinsic pathway, is critical in the initiation of fibrin formation. Coagulation factors is in two mechanisms, and in order to clot blood, they are activated by a cooperation with $Ca^{2+}$, phospholipid and vitamin K etc. For example, the human placental anticoagulant protein (PAP of PAP- I), which is a $Ca^{2+}$ -dependent phospholipid binding protein (Funakoshi et al., 1987) inhibited the activity of factor Xa, so that it prolonged fibrin formation. We wondered whether any other protein was involved in regulation of the coagulant system as an anticoagulant protein from natural organisms. Natural agents would have not harmful side-effects in comparision with chemically synthesized materials such as warfarin, aspirin, phenindione, etc.. But anticoagulant agents from natural, especially marine organisms have hardly been researched except for polysaccharides from marine algae. (omitted)

• The Korean Journal of Physiology and Pharmacology
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• v.12 no.5
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• pp.231-236
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• 2008

Heparin is a well-known anticoagulant widely used in various clinical settings. Interestingly, recent studies have indicated that heparin also has anti-inflammatory effects on neuroinflammation-related diseases, such as Alzheimer's disease and meningitis. However, the underlying mechanism of its actions remains unclear. In the present study, we examined the anti-inflammatory mechanism of heparin in cultured cerebral endothelial cells (CECs), and found that heparin inhibited the tumor necrosis factor $\alpha$ ($TNF{\alpha}$)-induced and nuclear factor kappa B (NF-${\kappa}B$)-dependent expression of adhesion molecules, such as intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1), which are crucial for inflammatory responses. Heparin selectively interfered with NF-${\kappa}B$ DNA-binding activity in the nucleus, which is stimulated by $TNF{\alpha}$. In addition, non-anticoagulant 2,3-O desulfated heparin (ODS) prevented NF-${\kappa}B$ activation by $TNF{\alpha}$, suggesting that the anti-inflammatory mechanism of heparin action in CECs lies in heparin's ability to inhibit the expression of cell adhesion molecules, as opposed to its anticoagulant actions.

• Journal of Microbiology and Biotechnology
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• v.18 no.3
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• pp.503-511
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• 2008

An edible marine red alga, Grateloupia filicina, collected from Jeju Island of Korea was hydrolyzed by cheap food-grade carbohydrases (Viscozyme, Celuclast, AMC, Termamyl, and Ultraflo) to investigate its anticoagulant activity. Among the tested enzymatic extracts of G. filicina, a Termamyl extract showed the highest anticoagulant activity. Anion-exchange chromatography on DEAE-cellulose and gel-permeation chromatography on Sepharose-4B were used to purify the active polysaccharide from the crude polysaccharide fraction of G. filicina. The purified sulfated polysaccharide (0.42 sulfate/total sugar) showed ${\sim}1,357kDa$ molecular mass and was comprised mainly of galactose(98%) and 1-2% of glucose. The sample showed potential anticoagulant activity on activated partial thromboplastin time (APTT) thrombin time (TT) assays. The purified G. filicina anticoagulant (GFA) inhibited the coagulation factor X (92%), factor II (82%), and factor VII (68%) of the coagulation cascade, and the molecular interaction (protein-polysaccharide) was highly enhanced in the presence of ATIII (antithrombin III). The dissociation constant of polysaccharide towards serine proteins decreased in the order of FXa (58.9 nM) >FIIa (74.6 nM) >FVII (109.3 nM). The low/less cytotoxicity of the polysaccharide benefits its use in the pharmaceutical industry; however, further studies that would help us to elucidate the mechanism of its activity are needed.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.2
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• pp.534-537
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• 2004

We produced twelve monoclonal antibodies(mAbs) against thyroglobulin and characterized the bindig profiles. Among them, three mAbs(TN-1, TN-2 and TN-3) were further characterized their binding specificities. TN-2 had a potent lupus anticoagulant activity and potentiated the anticoagulant effect of venom phospholipase A₂. he anticoagulant mechanism of TN-2 was elongation of the partial thromboplastin time and binding to phosphatidylserine which may have a pivot role in blood coagulation. And TN-2 was cross-reacted with ss-DNA and ds-DNA and had a characteristic of autoantibody. These results suggest that TN-2 may provide a useful tool for studying the correlation between autoimmune thyroiditis and its therapeutic effect.

• Proceedings of the Korean Society of Life Science Conference
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• 2003.10a
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• pp.158-158
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• 2003

• Journal of Ginseng Research
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• v.44 no.5
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• pp.717-724
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• 2020

Background: Malignant arrhythmias require drug therapy. However, most of the currently available antiarrhythmic drugs have significant side effects. Ginsenoside Rg2 exhibits excellent cardioprotective effects and appears to be a promising candidate for cardiovascular drug development. So far, the oral toxicity and antiarrhythmic effects of Rg2 have not been evaluated. Methods: Acute oral toxicity of Rg2 was assessed by the Limit Test method in mice. Subchronic oral toxicity was determined by repeated dose 28-day toxicity study in rats. Antiarrhythmic activities of Rg2 were evaluated in calcium chloride-induced arrhythmic rats. Antiarrhythmic mechanism of Rg2 was investigated in arrhythmic rats and H9c2 cardiomyocytes. Results: The results of toxicity studies indicated that Rg2 exhibited no single-dose (10 g/kg) acute oral toxicity. And 28-day repeated dose treatment with Rg2 (1.75, 3.5 and 5 g/kg/d) demonstrated minimal, if any, subchronic toxicity. Serum biochemical examination showed that total cholesterol in the high-dose cohort was dramatically decreased, whereas prothrombin time was increased at Day 28, suggesting that Rg2 might regulate lipid metabolism and have a potential anticoagulant effect. Moreover, pretreatment with Rg2 showed antiarrhythmic effects on the rat model of calcium chloride induced arrhythmia, in terms of the reduced duration time, mortality, and incidence of malignant arrhythmias. The antiarrhythmic mechanism of Rg2 might be the inhibition of calcium influx through L-type calcium channels by suppressing the phosphorylation of Ca²⁺/calmodulin-dependent protein kinase II. Conclusion: Our findings support the development of Rg2 as a promising antiarrhythmic drug with fewer side effects for clinical use.

• Korean Journal of Veterinary Service
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• v.22 no.4
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• pp.377-383
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• 1999

Polymorphonuclear (PMN) leucocytes are fundamental importance to the body's defense mechanism and play a major role in the local and systemic reactions to infectious disease. Investigation of the physiological and pathological role of the various leucocyte subtypes in host defence mechanisms is dependent upon the isolation of adequate numbers of viable, pure leucocyte fractions. This report describes the separate frequency of PMN leucocytes both from buffy coat layer and from packed RBC layer when bovine peripheral blood was treated with various anti-coagulants such as acid-citrate-dextrose(ACD), ethyldiaminetetraacetic acid(EDTA), sodium citrate and heparin. The separate frequencies of PMN leucocytes from buffy coat layer was 60.4$\pm$9.6%(heparin), 56.8$\pm$11.8%(sodium citrate), 30.6$\pm$14.1%(ACD) and 6.2$\pm$3.7%(EDTA), in order. Those from packed RBC layer monitored with EDTA, ACD, sodium citrate and heparin was 85.0$\pm$4.7%, 84.3$\pm$5.5%, 83.8$\pm$6.5% and 76.3$\pm$7.7%, respectively. The Ficoll-hypaque(FH) density gradient method was used to remove a small part of lymphocytes and/or monocytes from leucocytes in packed RBC layer. With the result that it increased separate frequency of PMN leucocytes from EDTA(89.9$\pm$2.4%), ACD(89.5$\pm$3.6%), and sodium citrate(83.6$\pm$10.3%) than heparin(68.4$\pm$13.9%). These results indicate that the use of EDTA and ACD as anticoagulant Is suitable for the separation of PMN leucocytes from bovine peripheral blood, and that the FH density gradient method is able to increase the separate frequency of PMN leucocytes from packed RBC layer.

• Journal of Chest Surgery
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• v.23 no.2
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• pp.222-230
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• 1990

Protamine, a polycationic peptide extracted from fish, has been widely used for the reversal of anticoagulant action of heparin. However it may cause untoward circulatory side effects including hypotension and bradyarrhythmia. Nowadays, histamine and prostacyclin are regarded as one of the causative agents in the underlying mechanism of hemodynamic changes. To certify the possible role of histamine and prostacyclin, we observed simultaneous changes of the hemodynamic status, plasma concentration of thromboxane B, and circulating platelet count before and after intravenous injection of protamine. Experimental dogs, weighing 12-14kg, were divided into 2 groups; group A animals [n=10], were pretreated with indomethacin[2.5mg/kg] and group B animals[n=10] were pretreated with chlorpheniramine[0.5mg/kg] Heparin[3mg/kg] and protamine [3mg/kg] were administered sequentially in both groups. The results were as follows ; 1. The mean systemic arterial pressure was maintained well in groups A, whereas in group B it decreased from 165\ulcorner18mmHg to 138\ulcorner30mmHg[p<0.01] and 151\ulcorner21 mmHg[p<0.05] at 1 minute and 2 minutes after protamine injection. The mean pulmonary arterial pressure was not changed significantly in group A, whereas in group B it increased from 852 mmHg to 11\ulcorner3 mmHg[p<0.05], 11\ulcorner3 mmHg[p<0.05] and 10\ulcorner3 mmHg[p<0.05] at 1 minute, 3 minutes and 5 minutes after protamine injection. 2 The thromboxane B2 was not changed significantly in group A, whereas in group B it increased from 399\ulcorner401 \ulcornerg/ml to 744\ulcorner615 \ulcornerg/ml[p<0.05] and 814\ulcorner1070 \ulcornerg/ml [p<0.0 5] at 1 minute and 3 minutes after protamine injection without concomitant changes of pulmonary vascular resistance and pulmonary capillary wedge pressure. 3. The number of circulating platelet was not changed in group A, whereas in group B it decreased from 207100\ulcorner103600/\ulcornerl to 159700\ulcorner90900/\ulcornerl [p<0.05] at 1 minute after protamine injection, Although thromboxane B2 and platelet count were changed significantly after protamine injection, they did not cause the remarkable hemodynamic changes. Considering the above results, hemodynamic changes may be caused mainly by prostacyclin rather than thromboxane or platelet. Therefore, the pretreatment with cyclooxygenase inhibitor would be beneficial to prevent circulatory adverse effects of protamine for the patients undergoing cardiac surgery.