• Biomolecules & Therapeutics
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• v.18 no.2
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• pp.205-210
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• 2010

Several studies have shown that plant-derived polyphenols reduce cardiovascular accidents in high-risk patients and the inhibition of platelet function may be responsible for part of this benefit. Lindera obtusiloba is widely used in traditional herbal medicine for the treatment of cardiovascular and inflammatory diseases. Therefore, the antiplatelet and antithrombotic activities of Lindera obtusiloba Extracts (LOE) on in vitro platelet aggregation, radical scavenging activity and in vivo murine pulmonary thrombosis were examined. LOE was able to directly scavenge the stable DPPH radical in a concentration-dependent manner and its $IC_{50}$ value was 3.9 ${\pm}$ 0.1 ${\mu}g$/ml. LOE significantly inhibited collagen- and ADP-induced platelet aggregation in a concentration-dependent manner and its $IC_{50}$ value is 0.9 ${\pm}$ 0.1 mg/ml and 0.4 ${\pm}$ 0.1 mg/ml respectively. The inhibitory effect of LOE was comparable to aspirin ($IC_{50}$ values were 1.0 ${\pm}$ 0.5 and 1.0 ${\pm}$ 0.7 mg/ml, respectively). Furthermore, oral administration of LOE suppressed the death of mice with pulmonary thrombosis induced by intravenous injection of collagen plus epinephrine. Taken together, our results suggest LOE may be a promising candidate for antithrombotic agent, and the antithrombotic effect of LOE may be due to, at least in part, antiplatelet activity.

• The Journal of Korean Obstetrics and Gynecology
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• v.18 no.3
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• pp.110-126
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• 2005

Purpose : The Purpose of this research was to investigate the effects of antithrombotic activities of Wuslsan (WSS). Methods : Measure the effect which was given to blood flow rate through the regular volume of glass tube after the blood was diluted five times with ACD soulution. 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 is caused from the administration of platelet aggregation regent. Being classified one group of eight mice, each of them was divided into Normal, Control, and WSS. The normal group supplied a saline solution and the control group brought the dextran extravasated blood after an hour of administering the saline solution. Also WSS was dissolved in 2ml saline solution and then we dosed it to the experimental mice with Oral Zonde one day before the experiment. After that, the mice were abstained from food. And then we gave a measured amount of it before an hour. Finally, it gave rise to dextran extravasated blood in the same way as the Control group. Results : The results were obtained as follows. WSS inhibited platelet aggregation induced by ADP and epinephrine significantly as compared with the control group. WSS showed fibrinolytic activity insignificantly as compared with the control group. WSS increased blood flow rate significantly as compared with the control group in vitro. WSS inhibited pulmonary embolism induced by collagen and Epinephrine(inhibitive rate is 37.5%). WSS increased number of platelet and fibrinogen amount significantly, and shortened PT and APTT as compared with the control group in thrombus model induced by dextran. Conclusion : WSS is effective antithrombotic activity from experimental result.

• The Journal of Korean Obstetrics and Gynecology
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• v.18 no.1
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• pp.1-14
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• 2005

This study was performed to evaluate antithrombotic activities of Saegeumsan (瑞金散, SGS) which has effects of activating blood, removing thrombus. This study is designed to measure the effect which was given to 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 is caused from the administration of platelet aggregation regent. We standardized the time when the experimental animals were incapable of functioning the hind legs more than 20 minutes or maintained trembling. Being classified one group of eight mice, each of them was divided into Normal, Control, and SGS. The normal group supplied a saline solution and the control group brought the dextran extravasated blood after an hour of administering the saline solution. Also, SGS was dissolved in $2m{\ell}$ saline solution and then we dosed it to the experimental mice with Oral Zonde one day before the experiment. After that, the mice were abstained from food. And then we gave a measured amount of it before an hour. Finally, it gave rise to dextran extravasated blood in the same way as the Control group. The results were obtained as follows, SGS significantly inhibited platelet aggregation induced by ADP and epinephrine when analyzed by the Sigmoid $E_{max}$ model in WinNonlin. $EC_{50}$ values of SGS were 4.61 mg/ml and 12.41 mg/ml for ADP and epinephrine respectively. SGS showed fibrinolytic activity insignificantly as compared with the control group. SGS increased blood flow rate significantly as compared with the control group in vitro. SGS inhibited pulmonary embolism induced by collagen and epinephrine(inhibitive rate is 37.5 %). SGS increased number of platelet and fibrinogen amount significantly, and shortened PT and APTT as compared with the control group in thrombus model induced by dextran. According to, SGS is effective antithrombotic activity from experimental result.

• The Journal of Korean Medicine
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• v.30 no.6
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• pp.1-8
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• 2009

Objectives: Formulas for treatment of yin or yang deficiency conditions have been commonly used in traditional Korean medicine. The aim of this study is to examine the possible inhibitory effects of yin- or yang-tonifying formulas on in vivo anti-platelet activity and in vivo anti-thrombotic activity. Methods: We tested the effects of 26 types of yin- or yang-tonifying formulas on platelet aggregation induced by collagen in human whole blood using the impedance method of aggregometry and accessed a biomarker of platelet activation using thromboxane $B_2$ immunoassay. We also tested the anti-thrombotic effects of effective candidates on experimental models of thrombosis in mice. Results: 3 types of yin-tonifying formulas and 3 types of yin-yang-tonifying formulas were selected to be the most effective candidates (p<0.01). Also, through in vivo study, the antithrombotic activities of Igyeong-tang, Gamisipjeondaebo-tang, and Gamisoyo-san-treated groups, with recovery rate of 60, 50, and 45.45%, respectively, were observed to be higher than those of the control group (saline, 36.8%) in mouse acute thrombosis. Conclusion: These results show that yin-tonifying formulas are more effective in anti-platelet and anti-thrombotic activity than yang-tonifying formulas.

• 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.

• Korean Journal of Microbiology
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• v.43 no.4
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• pp.298-303
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• 2007

Antithrombotic activities of water extract of cheongkookjang and cheongkookjang fermented with green tea or mugwort were evaluated on some antithrombosis related activities in vitro and thrombotic death inhibition in vivo. Cheongkookjang made of white soybean (Glycine max) or black small soybean (Rhynchosia nulubilis) showed potent antioxidative activities. Addition of green tea or mugwort during cheongkookjang fermentation increased the antioxidative activity, cheongkookjang with green tea showed more drastic increase compared with cheongkookjang with mugwort. Nitrite scavenging effects of the cheongkookjang extracts were prominent but the addition of green tea or mugwort seldom increased the scavenging effects. All the cheongkookjang extracts showed strong inhibitory activities on platelet aggregation. The inhibitory activities of cheongkookjang were increased considerably by addition of green tea or mugwort even with low concentration. Plasmin unit as fibrinolytic activity was not affected considerably by addition of green tea. Addition of mugwort decreased the activity transiently at low concentration ($0.3{\sim}1.0%$) but increased again slowly at higher concentration ($1{\sim}3%$). In vitro thrombotic death inhibition test, the antithrombotic activity of cheongkookjang made of black small bean with green tea was higher by about 1.5 times compared to that without green tea. As results, cheongkookjang might inhibit antithrombosis not only by fibrinolytic action but also by inhibition of platelet aggregation and antioxidative action. The addition of functional materials such as green tea or mugwort could increase the antithrombotic function, even at low concentration.

• Journal of the Korean Applied Science and Technology
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• v.19 no.3
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• pp.181-188
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• 2002

The purpose of this study was to determine the effect of dietary ${\gamma}$-linolenic acid on plasma lipid metabolism and anti thrombotic activity in male Sprague Dwaley Strain rats. Rats weighing an average of $100{\sim}120g$ were fed a experimental diets containing 5% lard (saturated fatty acids), corn oil(linoleic acid), evening promise oil(EPO, 9% ${\gamma}$-linolenic acid) or borage oil(BO, 24% ${\gamma}$-linolenic acid) for 3Odays, respectively. Though there were no significant difference in the food intake among the groups, the body weight gain of the BO group was significantly lower than that of other group. The spleen weight of the lard group was significantly lower than that of other group. The bleeding time of the BO group was significantly longer than that of other group. The blood clotting time was significantly tended to long in EPO and BO groups compared with lard group. The plasma triacylglyceride and total cholesterol concentration were high in order of lard, com oil, EPO and BO, groups and there were significant differences among the groups. The plasma HDL-C concentrations were high in order of BO, EPO, com oil and lard groups and there were significant differences among the groups. The plasma LDL-C concentrations were significantly the highest in lard group, but the lowest in BO group. These data indicate that ${\gamma}$-linolenic acid has a antithrombotic activity, and decrease the plasma triacylglyceride, total cholesterol and LDL-C concentrations in rats.

• Proceedings of the KOSOMBE Conference
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• v.1997 no.05
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• pp.51-54
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• 1997

Lumbrokinase, potent fibrinolytic enzyme purified from earthworm, was immobilized onto the total artificial heart valves using photoreaction. This valve were implanted into the lamb for three days. After experiments, thrombus was observed in the untreated valves whereas no thrombus was observed in the lumbrokinase immobilized valves. The fibrinolytic activity and proteolytic activity of the implanted valve was examined. The fibrinolytic activity of the valve was remained after the implantation. The lumbrokinase could be a suitable fibrinolytic agents in the vascular contacting devices to reduce the thrombus.

• Biomolecules & Therapeutics
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• v.7 no.3
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• pp.221-226
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• 1999

In this study, 75% ethanol extract from the flowers of Carthamus tinctorius was prepared and biological activities were examined. The extract showed the inhibitory activity of vascular smooth muscle contraction and antithrombotic activity judged by bleeding time measurement. It also showed anti-inflammatory and potent analgesic activities in vivo. By oral administration of the extract, no acute toxicity was observed up to 5 g/kg in mice and rats. All these results strongly suggest that this extract may be beneficial for postmenopausal disorder by enhancing blood circulation.

• BMB Reports
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• v.30 no.1
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• pp.37-40
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• 1997

Earthworm extracts are known for anti-inflammatory, analgesic. antipyretic, and anticancer effects but can also influence blood circulation. It was previously shown that an earthworm, Lumbricus rubelius. contained a water-extractable anticoagulant which was a heat- and acid-stable molecule with hydrophilic property. In order to uncover the biochemical nature of this molecule, the anticoagulant was processed with various hydrolases such as trypsin, DNase, RNase. and lysozome. When the digested samples were analyzed with an in vitro coagulation test measuring activated partial thromboplastin time (APTT) and agarose gel electrophoresis, the anticoagulant proved to be a relatively homogeneous DNA fragment with relative molecular size around 72 base pairs. Interestingly, the activity was further stimulated with a trypsin digestion. RNA. on the other hand, did not prolong the APTT. It was also demonstrated that the DNA accelerated the antithrombin III (AT-III) inhibition of thrombin from $IC_{50}$ of 0.34 to 0.16 unit determined with S-2238 as a substrate, whereas heparin, a popular anticoagulant. shifted the value to 0.05. Therefore, it is suggested that the DNA could be considered as an alternative antithrombotic agent to heparin, which would exhibits bleeding side effects.