• Applied Biological Chemistry
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• 제41권8호
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• pp.613-615
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• 1998

Antithrombotic function of doenjang was investigated using thoroughly desalted aqueous extract. In the presence of the extract, thrombin suffered loss of its fibrin-clotting activity, whose extent increased in a dose-dependent manner. The active substance involved in thrombin inhibition showed a high thermal stability. Results of Sephadex G-25 permeation chromatography indicated that there were at least two soluble thrombin inhibitors in doenjang with different molecular sizes.

• The Korean Journal of Physiology and Pharmacology
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• 제15권4호
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• pp.211-216
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• 2011

Glioblastoma multiforme is one of the most common and aggressive tumors in central nervous system. It often possesses characteristic necrotic lesions with hemorrhages, which increase the chances of exposure to thrombin. Thrombin has been known as a regulator of MMP-9 expression and cancer cell migration. However, the effects of thrombin on glioma cells have not been clearly understood. In the present study, influences of thrombin on glioma cell migration were examined using Boyden chamber migration assay and thrombin-induced changes in MMP-9 expression were measured using zymography, semi-quantitative RT-PCR, and Western blotting. Furthermore, underlying signaling pathways by which thrombin induces MMP-9 expression were examined. Thrombin-induced migration and MMP-9 expression were significantly potentiated in the presence of wortmannin, a PI3K inhibitor, whereas MAPK inhibitors suppressed thrombin-induced migration and MMP-9 expression in C6 glioma cells. The present data strongly demonstrate that MAPK and PI3K pathways evidently regulate thrombin-induced migration and MMP-9 expression of C6 glioma cells. Therefore, the control of these pathways might be a beneficial therapeutic strategy for treatment of invasive glioblastoma multiforme.

• 대한의생명과학회지
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• 제24권3호
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• pp.253-262
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• 2018

Platelets are activated at sites of vascular injury via several molecules, such as adenosine diphosphate, collagen and thrombin. Full platelet aggregation is absolutely essential for normal hemostasis. Moreover, this physiological event can trigger circulatory disorders, such as thrombosis, atherosclerosis, and cardiovascular disease. Therefore, platelet function inhibition is a promising approach in preventing platelet-mediated circulatory disease. Many studies reported the involvement of mitogen-activated protein kinases (MAPKs) signaling pathways in platelet functions. However, these studies were limited. Thus, we examined MAPK signaling pathways in human platelets using specific MAPK inhibitors, such as PD98059, SB203580, and SP600125. We observed that these inhibitors were involved in calcium mobilization and influx in human platelets. They also suppressed thrombin-induced ${\alpha}$- and ${\delta}$-granule release. These results suggest that PD98059, SB203580, and SP600125 exhibit $Ca^{2+}$ antagonistic effects.

• 생약학회지
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• 제35권1호통권136호
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• pp.52-61
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• 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.

• 생명과학회지
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• 제18권3호
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• pp.363-368
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• 2008

국내의 자생곤충 76종으로부터 다양한 용매를 이용하여 304종의 추출물을 조제한 후, 이들의 항혈전 활성을 평가하였다. 먼저 트롬빈 저해활성 평가 결과, 방아깨비, 왕잠자리, 비단노린재 및 끝검은메뚜기의 DMSO 추출물 4종, 알락수염노린재, 가시길쭉바구미, 잔날개여치, 털두꺼비하늘소, 송장벌레과 유충 및 등빨간거위벌레의 물 추출물 6종에서 우수한 활성을 확인하였다. 선정된 10종을 대상으로 aPTT 측정결과 비단노린재, 방아깨비 및 잔날개여치 추출물에서 미약한 증가가 나타났으나, 전반적으로 혈전 생성 저해의 유의적인 변화는 인정되지 않아, 이들 추출물들은 트롬빈을 직접 저해하는 것으로 추측되었다. 한편 항트롬빈 활성이 우수한 10종 추출물 중에서 방아깨비, 가시길쭉바구미, 알락수염노린재 및 송장벌레과 유충 추출물에서는 우수한 DPPH 소거능을 나타내어 이들의 약용 곤충자원으로의 개발 가능성을 확인하였다.

• BMB Reports
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• 제33권2호
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• pp.112-119
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• 2000

Three kinds of serine protease inhibitors, members of the Bowman-Birk trypsin inhibitor, were purified from Dolichos lablab seeds and named Dolichos protease inhibitor 1, 2 and 3 (DI-1, DI-2 and DI-3), respectively. Each inhibitor showed a single band with gel mobility at around 15.9, 12.1 and 14.6 kDa on 20% SDS-PAGE under reducing conditions. To characterize inhibitory specificity, the inhibition constant (Ki) for these inhibitors was measured against several known serine proteases. All three Dolichos protease inhibitors (DI-1, DI-2 and DI-3) inhibited the activity of trypsin and plasmin, but had no effect on thrombin and kallikrein (either for human plasma kallikrein or for porcine pancreas kallikrein). DI-1 inhibited chymotrypsin most effectively (Ki = $3.6{\times}10^{-9}\;M$), while DI-2 displayed inhibitory activity for porcine pancreatic elastase (Ki = $6.2{\times}10^{-8}\;M$). Pre-treatment of the 33 mg/kg of DI-mixture (active fractions from $C_{18}$ open column chromatography that included DI-1, DI-2 and DI-3) inhibited the induction of pseudomonal elastase-induced septic hypotension and prevented an increase in bradykinin generation in pseudomonal elastase-treated guinea pig plasma. Also, the increase of kallikrein activity, by injection of pseudomonal elastase, was inhibited by the pretreatment of the DI-mixture in a guinea pig. Since the DI-mixture had no inhibitory effect on kallikrein activity when Z-Phe-Arg-MCA was used as a substrate in vitro, its inhibitory activity in the pseudomonal elastase-induced septic hypotension model might not be due to a direct inhibition of plasma kallikrein in the activation cascade of the Hageman factor and prekallikrein system. These results suggest that the Dolichos DI-mixture might be used as an inhibitor in pathogenic bacterial protease-induced septic shock.

• 생명과학회지
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• 제20권11호
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• pp.1640-1647
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• 2010

혈전성 질환을 예방, 개선할 수 있는 안전한 항혈전제를 개발하기 위해, 일반적으로 항혈전 효과가 있다고 알려진 식용 해조류 35종(갈조류 17종, 홍조류 11종 및 녹조류 7종)을 대상으로 메탄올 추출물을 조제하고, 이들의 thrombin time (TT), prothrombin time (PT), activated partial thromboplastin time (aPTT)을 평가하였다. 트롬빈 저해(TT)의 경우 35종 중에서 감태, 곰피, 대황, 넓패, 패, 알송이모자반 및 야마다모자반에서 강력한 활성을, 프로트롬빈 저해(PT)의 경우 트롬빈 저해활성이 나타난 7종 중 야마다모자반을 제외한 6종에서 강력한 활성을 확인하였으며, 혈액응고인자 저해(aPTT)에서는 감태, 곰피, 대황, 넓패, 패, 알송이모자반 및 톳에서 강력한 활성이 나타났다. 선별된 8종의 시료를 대상으로 다양한 농도에서 항혈전 활성을 평가한 결과 알송이모자반에서 가장 강력한 활성을 확인하였으며, 곰피, 대황 및 넓패가 효과적이었다. 항혈전 활성과 해조류 추출물의 총 flavonoid, 총 polyphenol, 총당 및 환원당 함량과의 상관 관계를 검토한 결과, 활성물질은 flavonoid성 물질로 추측되었으며, 선별된 8종은 총 flavonoid 함량 및 총 polyphenol 함량이 매우 높은 특징을 나타내었다. 본 연구 결과는 경제적이면서 대량공급이 가능한 식용 해조류로부터 신규의 안전한 항혈전제 개발이 가능함을 제시하며, 야마다모자반과 톳은 트롬빈 또는 혈장내 응고 인자에 대한 특이 저해제로 개발 가능함을 제시하고 있다.

• Bulletin of the Korean Chemical Society
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• 제35권6호
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• pp.1763-1768
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• 2014

In the course of our program to search for antithrombotic and anti-inflammatory agents from plants, twelve phenolics (1-12) were isolated from the stems of Parthenocissus tricuspidata. Their structures were elucidated on the basis of spectroscopic (1D and 2D NMR, and MS) data analyses, and comparison with published data. At the concentration of $100{\mu}g/ml$, compounds 2, 4, 6 and 10 possessed potential effects on anti-blood coagulation, with inhibitory percentage of 216, 174, 148 and 225%, respectively; while aspirin used as positive control showed 181% inhibition at the same concentration. Furthermore, the anti-inflammatory activity of isolated compounds (1-12) was investigated on lipopolysaccharide (LPS)-induced murine macrophage cells (RAW264.7). Compounds 2, 4 and 6 also potential inhibited the production of nitric oxide, with $IC_{50}$ values of $11.9{\pm}0.3$, $2.9{\pm}0.2$ and $29.0{\pm}0.6{\mu}M$, respectively. Celastrol, the positive control used, gave an $IC_{50}$ value of $1.0{\pm}0.1{\mu}M$.

• Archives of Pharmacal Research
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• 제27권6호
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• pp.619-623
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• 2004

Tissue factor (TF, tissue thromboplastin) is a membrane bound glycoprotein, which acceler-ates the blood clotting, activating both the intrinsic and the extrinsic pathways to serve as a cofactor for activated factor VII (Vila). The TF-factor Vila complex (TF/VIIa) proteolytically activates factors IX and X, which leads to the generation of thrombin and fibrin clots. In order to isolate TF inhibitors, by means of a bioassay-directed chromatographic separation technique, from the leaves of Eriobotrya japonica Lindley (Rosaceae), a known sesquiterpene glycoside (2) and ferulic acid (3) were isolated as inhibitors that were evaluated using a single-clotting assay method for determining TF activity. Another sesquiterpene glycoside (1) was also isolated but was inactive in the assay system. Compound 3 was yielded by alkaline hydrolysis of compound 2. The structures of compounds 1, 2, and 3 were identified by means of spectral analysis as $3-O-{\alph}-L-rhamnopyranosyl-(1{\rightarrow}4)-a-L-rhamnopyranosyl-(1{\rightarrow}2)-[{\alph}-L-rhamnopyrano-syl-(1{\rightarrow}6)]-{\beta}-D-glucopyranosyl nerolidol$ (1), $3-O-{\alph}-L-rhamnopyranosyl-(1{\rightarrow}4)-{\alph}-L-rhamnopyr-anosyl-(1{\rightarrow}2)-[{\alph}-L-(4-trans-feruloyl)-rhamnopyranosyl-(1{\rightarrow}6)]-{\beta}-D-glucopyranosyl$ nerolidol (2) and ferulic acid (3), respectively. Compounds 2 and 3 inhibited 50% of the TF activity at con-centrations of 2 and $369{\;}\mu\textrm{m}/TF$ units, respectively.

• 한국미생물·생명공학회지
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• 제51권2호
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• pp.167-173
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• 2023

Proteolytic enzymes secreted by Aspergillus, as pathogenicity factors, affect blood coagulation and fibrinolysis, and therefore the target proteins of their action in the bloodstream are of significant interest. In the present study, the action of the isolated protease of A. terreus 2 on different human plasma proteins was shown. The protease of A. terreus 2 exhibited the highest proteolytic activity against hemoglobin, which was 2.5 times higher than the albuminolytic activity shown in both of the protein substrates used. In addition, the protease has significant ability to hydrolyze both fibrin and fibrinogen. However, the inability of the A. terreus 2 protease to coagulate rabbit blood plasma and coagulate human and bovine fibrinogen indicates the severity of the enzyme's action on human blood coagulation factors. It should be considered as a potential indicator of this isolated protease's participation in fungal pathogenesis. The protease shows no hemolytic activity. Furthermore, its activity is insignificantly inhibited by thrombin inhibitors, and is not inhibited by plasmin inhibitors.