• Applied Biological Chemistry
• /
• v.41 no.8
• /
• pp.613-615
• /
• 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
• /
• v.15 no.4
• /
• pp.211-216
• /
• 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.

• Biomedical Science Letters
• /
• v.24 no.3
• /
• pp.253-262
• /
• 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.

• Korean Journal of Pharmacognosy
• /
• v.35 no.1 s.136
• /
• 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.

• Journal of Life Science
• /
• v.18 no.3
• /
• pp.363-368
• /
• 2008

The in vitro thrombin inhibitory activities of 304 crude extracts from 76 kinds of korean wild insects were evaluated. Measurement of thrombin time showed that the DMSO extracts of Acrida cinerea cinerea (Thunberg), Anax parthenope julius Brauer, Eurydema rugosa Motschulsky, and Stethophyma magister (Rehn) and the water extracts of Dolycoris baccarum Linne, Lixus divaricatus Motschulsky, Metrioptera bonneti, Moechotypa diphysis (Pascoe), Nicrophorus concolor sp., and Tomapoderus ruficollis (Fabricius) had strong thrombin inhibitory activity. No prominent changes of activated partial thromboplastin time by treatment of the selected extracts suggested direct inhibition of thrombin activity by the insect extracts. DPPH scavenging activity of selected extracts showed that the extract of A. cinerea cinerea (Thunberg), D. baccarum Linne, L. divaricatus Motschulsky and N. concolor sp. has good antioxidant activity as well as antithombin activity. Our results suggested that some of korean wild insects could be developed as a natural source of antithrombosis.

• BMB Reports
• /
• v.33 no.2
• /
• pp.112-119
• /
• 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.

• Journal of Life Science
• /
• v.20 no.11
• /
• pp.1640-1647
• /
• 2010

Seaweeds have been recognized as a health food, having anti-obesity, anti-constipation and anticoagulation activities, and the use of seaweeds in the food, medicine, and cosmetic industries have recently significantly increased. In this study, methanol extracts were prepared from 35 different seaweeds (17 phaeophyta, 11 rhodophyta and 7 chlorophyta), and thrombin time (TT), prothrombin time (PT) and activated partial thromboplastin time (aPTT) were determined in order to develop safe and novel anticoagulation agents from natural products. In TT experiments, Ecklonia cava, Ecklonia stolonifera, Eisenia bicyclis (Kjellman) Setchell, Ishige foliacea, I. okamurai, Sargassum confusum and S. yamade showed strong thrombin inhibition activity among the 35 different seaweeds. In PT experiments, the inhibitions of prothrombin were identified in the selected seaweeds from TT experiment, with the exception of S. yamade. In aPTT experiments, the seaweeds with blood coagulation inhibition factors were E. cava, E. stolonifera, E. bicyclis (Kjellman) Setchell, I. foliacea, I. okamurai, S. confusum and Hixikia fusiforme Okamura. Further anticoagulation assay with the selected 8 seaweeds suggested that S. confusum is most effective in antithrombosis, and E. stolonifera, E. bicyclis (Kjellman) Setchell, and I. foliacea have high potential as antithrombosis agents. Based on components-activity correlation analysis, flavonoids are considered as active anticoagulation components of seaweeds These results suggest that edible seaweeds, especially S. confusum, have potential as safe and novel anticoagulants, and S. yamade and H. fusiforme Okamura could be used as a thrombin-specific and coagulation factor-specific inhibitors.

• Bulletin of the Korean Chemical Society
• /
• v.35 no.6
• /
• pp.1763-1768
• /
• 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
• /
• v.27 no.6
• /
• pp.619-623
• /
• 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.

• Microbiology and Biotechnology Letters
• /
• v.51 no.2
• /
• pp.167-173
• /
• 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.