• KSBB Journal
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• 제25권1호
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• pp.11-17
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• 2010

본 논문에서는 혈관내피세포에서 저농도의 트롬빈이 TNF-$\alpha$가 NF-kB의 활성화를 통해 생성되는 IL-6의 생성량에 미치는 영향을 관찰하였다. TNF-$\alpha$는 혈관내피세포에서 NF-kB의 활성화를 통해 염증을 유발시킨다는 것은 잘 알려진 사실이다. 이 논문에서는 TNF-$\alpha$가 매개하는 염증작용에서 저농도의 트롬빈은 TNF-$\alpha$가 생성시키는 IL-6의 생성량을 감소시켰고, 여기에는 트롬빈의 수용체인 PAR-1이 작용하다는 것을 확인하였다. 뿐만 아니라, 세포내의 PI3-Kinase 역시 저농도 트롬빈이 관여한다는 것을 확인하였다. 이것은 저농도의 트롬빈이 수용체인 PAR-1을 활성화시키고, 활성화된 PAR-1 은 PI3-Kinase의 활성화을 통해 항염증작용을 보여준디는 것을 의미한다. 이 결과는 향후 중증 패혈증 및 각종 염증질환을 치료할 수 있는 신약개발에 있어 중요한 단서를 제공하고 혈관내피세포에서 아직 명확하게 밝혀지지 않은 트롬빈의 염증작용 및 항염증작용의 기전을 밝히는데 좋은 정보를 제공할 것이다.

• 대한의생명과학회지
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• 제16권4호
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• pp.271-277
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• 2010

This study was conducted to investigate the biological activities of Areca catechu L. The antioxidative, fibrinolytic, thrombin inhibitory, and ${\alpha}$-glucosidase inhibitory activities of Areca catechu L extracted with hexane, $CHCl_3$, ethyl acetate, butanol, and water were measured. The water fraction showed the highest extraction yield at 3.65% (w/w). The butanol, $CHCl_3$, water, and ethyl acetate fractions showed strong antioxidative activities at 81.6%, 87.1%, 88.0%, and 89.5%, respectively. The fibrinolytic activity was strong only in the ethyl acetate fraction at 0.84 plasmin units/ml. The 100-fold dilution of the water fraction had the strongest thrombin inhibitory activity at 59.2%. The 100-fold dilution of butanol fraction displayed the strongest ${\alpha}$-glucosidase inhibitory activity at 88.6%. In conclusion, the extracts of Areca catechu L hold promise for use in the development of biofunctional foods to prevent cardiovascular diseases.

• 생명과학회지
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• 제24권8호
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• pp.873-879
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• 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.

• BMB Reports
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• 제46권11호
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• pp.544-549
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• 2013

High mobility group box 1 (HMGB1) is involved in the pathogenesis of vascular diseases. Unlike activated protein C (APC), the activation of PAR-1 by thrombin is known to elicit proinflammatory responses. To determine whether the occupancy of EPCR by the Gla-domain of APC is responsible for the PAR-1-dependent antiinflammatory activity of the protease, we pretreated HUVECs with the PC zymogen and then activated PAR-1 with thrombin. It was found that thrombin downregulates the HMGB1-mediated induction of both TNF-${\alpha}$ and IL-6 and inhibits the activation of both p38 MAPK and NF-${\kappa}B$ in HUVECs pretreated with PC. Furthermore, thrombin inhibited HMGB1-mediated hyperpermeability and leukocyte adhesion/migration by inhibiting the expression of cell adhesion molecules in HUVECs if EPCR was occupied. Collectively, these results suggest the concept that thrombin can initiate proinflammatory responses in vascular endothelial cells through the activation of PAR-1 may not hold true for normal vessels expressing EPCR under in vivo conditions.

• 대한의생명과학회지
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• 제14권4호
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• pp.249-255
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• 2008

In this study we investigated the biological activities of Crataegi Fructus, including antioxidative, fibrinolytic, $\alpha$-glucosidase inhibitory, and thrombin inhibitory activities. Crataegi Fructus, hot water extract was fractionated into hexane, $CHCLl_3$, ethyl acetate, butanol, and water fractions, and each of these was assayed individually. The water fraction showed the highest extraction yield at 4.08% (w/w). The antioxidative activities of the water, ethyl acetate, and butanol fractions were 31.07%, 45.87%, 50.28%, and 91.74%, respectively. Assays for fibrinolytic activity indicated that only the butanol fraction has significant efficacy at 1.93 plasmin units/ml. Thrombin inhibitory assays indicated that the 10-fold dilutions of the $CHCLl_3$, ethyl acetate, and butanol fractions had inhibitory activities of 34.97%, 41.43%, and 58.10%, respectively. The 10-fold dilutions of the only ethyl acetate fraction demonstrated $\alpha$-glucosidase inhibitory activities of 75.07%. From the above results, we propose that extracts of Crataegi Fructus can be used as a material for the development of biofunctional foods.

• BMB Reports
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• 제46권10호
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• pp.484-489
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• 2013

Piperlonguminine (PL), an important component of Piper longum fruits, is known to exhibit anti-hyperlipidemic, antiplatelet and anti-melanogenic activities. Here, the anticoagulant activities of PL were examined by monitoring activated-partial-thromboplastin-time (aPTT), prothrombin-time (PT), and the activities of thrombin and activated factor X (FXa). The effects of PL on the expressions of plasminogen activator inhibitor type 1 (PAI-1) and tissue-type plasminogen activator (t-PA) were also tested in tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) activated HUVECs. The results showed that PL prolonged aPTT and PT significantly and inhibited the activities of thrombin and FXa. PL inhibited the generation of thrombin and FXa in HUVECs. In accordance with these anticoagulant activities, PL prolonged in vivo bleeding time and inhibited TNF-${\alpha}$ induced PAI-1 production. Furthermore, PAI-1/t-PA ratio was significantly decreased by PL. Collectively, our results suggest that PL possesses antithrombotic activities and that the current study could provide bases for the development of new anticoagulant agents.

• Journal of Ginseng Research
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• 제40권1호
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• pp.76-85
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• 2016

Background: Binding of adhesive proteins (i.e., fibrinogen, fibronectin, vitronectin) to platelet integrin glycoprotein IIb/IIIa (${\alpha}IIb/{\beta}3$) by various agonists (thrombin, collagen, adenosine diphosphate) involve in strength of thrombus. This study was carried out to evaluate the antiplatelet effect of total saponin from Korean Red Ginseng (KRG-TS) by investigating whether KRG-TS inhibits thrombin-induced binding of fibrinogen and fibronectin to ${\alpha}IIb/{\beta}3$. Methods: We investigated the effect of KRG-TS on phosphorylation of vasodilator-stimulated phosphoprotein (VASP) and dephosphorylation of phosphatidylinositol 3-kinase (PI3K) and Akt, affecting binding of fibrinogen and fibronectin to ${\alpha}IIb/{\beta}3$, and clot retraction. Results: KRG-TS had an antiplatelet effect by inhibiting the binding of fibrinogen and fibronectin to ${\alpha}IIb/{\beta}3$ via phosphorylation of VASP ($Ser^{157}$), and dephosphorylation of PI3K and Akt on thrombin-induced platelet aggregation. Moreover, A-kinase inhibitor Rp-8-Br-cyclic adenosine monophosphates (cAMPs) reduced KRG-TS-increased VASP ($Ser^{157}$) phosphorylation, and increased KRG-TS-inhibited fibrinogen-, and fibronectin-binding to ${\alpha}IIb/{\beta}3$. These findings indicate that KRG-TS interferes with the binding of fibrinogen and fibronectin to ${\alpha}IIb/{\beta}3$ via cAMP-dependent phosphorylation of VASP ($Ser^{157}$). In addition, KRG-TS decreased the rate of clot retraction, reflecting inhibition of ${\alpha}IIb/{\beta}3$ activation. In this study, we clarified ginsenoside Ro (G-Ro) in KRG-TS inhibited thrombin-induced platelet aggregation via both inhibition of $[Ca^{2+}]_i$ mobilization and increase of cAMP production. Conclusion: These results strongly indicate that KRG-TS is a beneficial herbal substance inhibiting fibrinogen-, and fibronectin-binding to ${\alpha}IIb/{\beta}3$, and clot retraction, and may prevent platelet ${\alpha}IIb/{\beta}3$-mediated thrombotic disease. In addition, we demonstrate that G-Ro is a novel compound with antiplatelet characteristics of KRG-TS.

• 한국균학회지
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• 제42권3호
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• pp.207-212
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• 2014

검은비늘버섯을 이용하여 심혈관계 질환과 관련된 기능성 식품 개발을 위한 기초자료를 얻기 위해 검은비늘버섯물 추출물과 유기용매 분획물을 이용하여 심혈관계 질환관련 생리활성을 확인하였다. 물 추출물은 57.57%의 항산화활성을 나타냈지만 혈전용해활성, 트롬빈저해활성, ${\alpha}$-glucosidase 저해효과는 매우 낮게 나타났다. 반면 에틸 아세테이트 분획물은 0.70 plasmin unit의 높은 혈전용해 활성과, 87.36%와 46.44%의 높은 트롬빈 저해활성과 ${\alpha}$-glucosidase 저해활성을 나타냈으며, 물 분획물은 48.27%의 항산화활성을 나타냈다. 혈전용해활성이 높은 에틸 아세테이트 분획물은 섬유소원의 $A{\alpha}$와 $B{\beta}$ chain은 모두 분해시켰지만 ${\gamma}$ chain과는 반응하지 않았으며, $100^{\circ}C$에서 10분간 가열에도 혈전용해 활성에 변화가 없었다. 실험 결과로부터 검은비늘버섯의 에틸아세테이트 분획물은 높은 혈전용해활성과 트롬빈 저해활성과 ${\alpha}$-glucosidase 저해활성을 나타내 심혈관계 질환 관련 치료나 예방을 위한 기능성식품 개발에 이용 가능할 것으로 기대된다.

• 대한의생명과학회지
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• 제17권2호
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• pp.157-165
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• 2011

In this study, we investigated the biological activities of Carthamus tinctorius including antioxidative, fibrinolytic, thrombin inhibitory, ${\alpha}$-glucosidase inhibitory and collagenase inhibitory activities. Carthamus tinctorius, hot water extract was fractionated into hexane, chloroform, ethyl acetate, butanol and water fraction. Each of these was assayed individually. The hot water extract showed high antioxidative activity and thrombin inhibitory activity at 90.17% and 97.10% respectively. In the fraction activity tests, chloroform fraction showed the highest antioxidative activity at 81.85%. The fibrinolytic activity was strong only in the butanol fraction at 0.70 plasmin units/ml. The thrombin inhibitory activities of hexane, ethyl acetate and butanol fractions were 97.35%, 86.74% and 93.18% respectively. In collagenase inhibitory activity test, hexane fraction showed the highest activity at 87.78%. In conclusion, the hot water extract and solvent fractions of Carthamus tinctorius L can be used as a material for the development of biofunctional tea and foods respectively.

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