• Biomolecules & Therapeutics
• /
• 제1권2호
• /
• pp.166-170
• /
• 1993

Hirudin is a potent inhibitor of thrombin, which was originally obtained from the medicinal leech (Hirudo medicinalis) Now it is being produced through the recombinant technology on a large scale. Recombinant hirudin has been assayed for the anticoagulant activity by the measurement of clotting time and the inhibition of thrombin actvity using a chromogenic substrate. The assay range of partial thromboplastin time and thrombin time is within $0.2{\sim}1.0 {\mu}g/mι.$ Thrombin time is more sensitive to the measurement of clot. Ex vivo study showed the level of hirudin in rat plasma was highest in 10 min and then it was eliminated slowly. The half-life of r-hirudin was 80~110 min depending on the assay methods. Intraveneous injection of russel viper venom was used for thrombus induction combined with vents cava ligation. Inhibition of venous thrombosis was observed with i.v. hirudin. It was dependent on the concentration of hirudin.

• Journal of Ginseng Research
• /
• 제19권3호
• /
• pp.206-211
• /
• 1995

This study was investigated to find the effects of petroleum ether extract (Lipophilic fraction) from Korean red ginseng on platelet aggregation and thrombin time of the plasma in two groups of the experimental rats. One group of rats were fed with 15% corn oil (15%kg-diet) containing a number of 18 : 2 (linoleic acid) or 15% beef-tallow (15%/kg-diet) containing saturated fatty acids for 3 weeks, and were followed by feeding the petroleum ether extract (25 mg/kg-diet) for 3 weeks. The other group of rats (control group) were fed with 15% corn oil or 15% beef-tallow for 6 weeks. The platelet aggregation induced by thrombin and collagen was significantly inhibited and the thrombin time was prolonged in the 15% corn oil plus petroleum ether extract administrated group than in the 15% corn oil administrated group. And the same results were shown in the 15% beef-tallow plus petroleum ether extract administrated group. These results suggest that the petroleum ether extract from Korean red ginseng may have the beneficial effects on the inhibition of the platelet aggregation and the inhibition of blood coagulation induced by dietary fats.

• Archives of Pharmacal Research
• /
• 제28권6호
• /
• pp.667-674
• /
• 2005

The leaves of Persimmon (Diospyros kaki L.) has long been used for tea in Korea since it was thought to be effective against hypertension. An anticoagulant fraction was purified through gel filtration G-100, hydrophobic, gel filtration G-150, and FPLC, Phenyl superpose column chromatographies. The purified fraction was homogenous and its Mr was estimated 10,000 Da by gel filtration and SDS-PAGE. The purified fraction was sensitive to treatment of subtilisin B, but not to heat and its activity was not changed after periodate oxidation, indicating that the activity was not due to carbohydrates. It delayed thrombin time (TT), activated partial thromboplastin time (APTT), and prothrombin time (PT) using human plasma. TT was more sensitive than APTT and PT, suggesting that the anticoagulant activity may be caused by a degradation or a defect of fibrin or thrombin. It did not cause the hydrolysis of fibrin after incubation. However, it inhibited thrombin-catalyzed fibrin formation with a competitive inhibition pattern. These results indicate that it may be an antithrombotic agent and that it is bound to fibrinogen binding sites of thrombin.

• 생약학회지
• /
• 제35권1호통권136호
• /
• pp.52-61
• /
• 2004

Inhibitory activities of 264 methanol extracts, which were prepared from different parts of 210 kinds of wild and medicinal plants, against human thrombin were evaluated. Based on the anti-coagulation activity determined by thrombin time and activated partial thromboplastin time, the 14 extracts were screened. The fibrinolytic activity, heat stability and inhibition of other proteolytic digestive enzymes, such as pepsin, papain, trypsin and chymotrypsin, of the 14 extracts were further determined, and Ginko biloba (herba), Ephedra sinica (radix), Reynoutria elliptica (herba), Amomum tsao-ko Crevost (fructus), and Magnolia officinalis Rehd. et Wils (bark) were finally selected as possible plant sources for anti-thrombosis agent. These results suggested that medicinal and wild plants could be the potential source of thrombin inhibitor.

• Archives of Pharmacal Research
• /
• 제24권2호
• /
• pp.109-113
• /
• 2001

Curdlan is a natural $\beta$-1,3-glucan produced by Agrobacterium biovar 1. In this study, the anticoagulant activity of sulfoalkyl derivatives of curdlan was investigated by carrying out activated partial thromboplastin time (APTT) assay and compared with that of o-sulfonated curdlan. Approximately 100-fold higher concentration of o-sulfonated curdlan than heparin was required to obtain the same level of the clotting time. Anticoagulant activity of curdlan derivatives was dependent on the degree of sulfation in prolonging the clotting time. However, the chain length of the substituent did not play a role in prolonging the clotting time. The curdlan derivatives enhanced thrombin inhibition by mediating through antithrombin III. The inhibition of thrombin by o-sulfonated curdlan was found to be approximately 10-fold weaker than that by heparin.

• BMB Reports
• /
• 제47권7호
• /
• pp.376-381
• /
• 2014

Enzymatic oxidation of pyrogallol was efficiently transformed to an oxidative product, purpurogallin (PPG). Here, the anticoagulant activities of PPG were examined by monitoring activated partial thromboplastin time (aPTT), prothrombin time (PT), and the activities of thrombin and activated factor X (FXa). And, the effects of PPG on expression of plasminogen activator inhibitor type 1 (PAI-1) and tissue-type plasminogen activator (t-PA) were evaluated in tumor necrosis factor (TNF)-${\alpha}$ activated human umbilical vein endothelial cells (HUVECs). Treatment with PPG resulted in prolonged aPTT and PT and inhibition of the activities of thrombin and FXa, as well as inhibited production of thrombin and FXa in HUVECs. In addition, PPG inhibited thrombin-catalyzed fibrin polymerization and platelet aggregation. PPG also elicited anticoagulant effects in mice. In addition, treatment with PPG resulted in significant reduction of the PAI-1 to t-PA ratio. Collectively, PPG possesses antithrombotic activities and offers a basis for development of a novel anticoagulant.

• 생명과학회지
• /
• 제17권4호
• /
• pp.535-539
• /
• 2007

식용 및 약용으로 광범위하게 이용되고 있는 산사자의 유용 생리활성 검토를 목적으로 산사자 추출물 및 분획물을 조제하여, 혈액내의 혈전 생성에 직접 관여하는 인간 트롬빈의 thrombin time(TT) 및 activated partial thromboplastin time(aPTT) 증가 활성을 측정하여 항혈전 활성을 평가하였다. 산사자의 항혈전 물질은 물 추출보다는 메탄을 추출이 적합하며(추출효율 40.4%), butanol 분획에서 가장 강력한 활성을 나타내었다. 산사자의 butanol 분획물은 1.25 mg/ml의 농도에서 835%의 TT 증가, 315%의 aPTT 증가를 보여 아스피린보다 2.9배 이상의 강력한 항혈전 활성을 나타내었다. 이러한 항혈전 활성은 기존의 보고된 quercetin, kaempferol, myricetin 및 rutin과는 무관하였다. 또한 산사자의 활성물질은 0.5 N HCl 처리시 120분 이상 안정하였으나, $100^{\circ}C$, 30분 열처리에는 85% 이상의 활성소실이 나타나 열에 매우 민감한 물질로 확인되었다. 본 연구결과는, 비열 처리와 적합한 가공 공정이 가능하다면 산사자 추출물이 혈액순환 장해 및 혈관계 질환 예방 및 치료제로 개발 가능함을 제시하고 있다.

• 생명과학회지
• /
• 제14권3호
• /
• pp.509-513
• /
• 2004

혈전 관련 질환 예방 및 혈류개선 기능성 식품의 개발을 목표로, 식용 및 약용 산채류 55종의 메탄을 추출물을 조제한 후 트롬빈 저해활성을 검색하였다. 그 결과, 대조군으로 사용된 아스피린보다 우수한 항혈전 활성을 나타내는 붉나무, 사삼, 우산나물, 쥐다래, 참나물, 할미밀망, 큰까치수영, 첫버들 추출물 등 8종을 선별하였으며, aPTT, 혈전 용해활성, 열 안정성, 기타 소화효소 저해활성 등을 평가하며 참나물, 큰까치수영 및 갯버들 추출물을 최종 선별하였다. 참나물의 경우 열안정성이 낮은 반면 소화효소 비저해로 인해 그 이용성이 높으며, 큰 까치수영 및 갯버들의 경우 부분적인 소화효소 저해능이 있으나 우수한 항혈전활성 및 열 안정성을 나타내어 다양한 식품소재로 개발이 가능함을 확인하였다.

• BMB Reports
• /
• 제45권7호
• /
• pp.390-395
• /
• 2012

Oleanolic acid (OA), a triterpenoid known for its anti-inflammatory and anti-cancer properties, is commonly present in several medicinal plants but its anticoagulant activities have not been studied. Here, the anticoagulant properties of OA were determined by monitoring activated partial thromboplastin time (aPTT), prothrombin time (PT), fibrin polymerization as well as cell-based thrombin and activated factor X (FXa) generation activities. Data showed OA prolonged aPTT and PT significantly and inhibited thrombin catalyzed fibrin polymerization. In addition, OA inhibited the activities of thrombin and FXa and inhibited the generation of thrombin or FXa in human endothelial cells. OA also inhibited TNF-${\alpha}$-induced tissue factor expression on human endothelial cells. In accordance with these anticoagulant activities, OA showed an anticoagulant effect in vivo. These results indicate that OA possesses antithrombotic activities and suggest that daily consumption of a herb containing OA may be preventing thrombosis in pathological states.

• 생명과학회지
• /
• 제24권8호
• /
• pp.873-879
• /
• 2014

소나무(Pinus densiflora)의 잎의 성분을 분리하여 항트롬빈 활성을 조사하였다. 잎은 70% ethanol로 3회 추출하였으며, 그 추출물은 순차적으로 chloroform과 n-buthanol로 분획하였다. n-buthanol 분획으로부터 얻은 수용성 분획을 MPLC와 HPLC에 의해 분리하였다. 분리한 compound를 $^1H-$과 $^{13}C-NMR$로 동정한 결과 2, 3-dihydro-7-hydroxyl-3-hydroxymethyl-2-(4'-hydroxyl-3-methoxyphenyl)-5-benzofuranpropanol-3-O-${\alpha}$-rhamnopyranoside(a neolignan glycoside)와 2, 3-dihyro-3-hydroxymethyl-7-methoxy-2-(4'-hydroxyphenyl-3'-methoxy)-5-benxofuran propanol 4'-O-${\alpha}$-rhamnopyranoside (icariside $E_4$)의 neolignan 구조임을 확인하였다. 트롬빈 저해활성은 분리된 neolignan glycoside와 icariside $E_4$을 작용하여 혈장안에서 응고시간으로 측정하였다. 그 결과, neolignan glycoside의 응고시간이 icariside $E_4$보다 4배 이상 지연하였다. 저해활성은 neolignan glycoside의 농도가 증가함에 따라 그 시간이 지연되는 것을 확인하였다. 더 나아가 지연기전을 확인하기 위해 thrombin과 순수한 fibrinogen를 반응하였다. 그 결과, 트롬빈과 순수한 피브리노겐의 작용에 의해 응고시간은 지연되었으며, 이는 neolignan glycoside가 피브린형성에 주요한 트롬빈의 활성을 직접 저해하는 것으로 사료된다.