• BMB Reports
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• v.33 no.2
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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.

• Journal of Physiology & Pathology in Korean Medicine
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• v.25 no.3
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• pp.471-481
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• 2011

Clove has been frequently used as anti-diabetic, anti-microbial, anti-inflammatory, anesthetic drug and remedies for stomachache by coldness. In this study, the antioxidant activity of extract from Clove was studied in vitro methods by measuring the antioxidant activity by TEAC, measuring the scavenging effects on reactive oxygen species (ROS) [superoxide anion, hydroxyl radical] and on reactive nitrogen species (RNS) [nitric oxide and peroxynitrite] as well as measuring the inhibitory effect on $Cu^{2+}$-induced human LDL oxidation. Anti-platelet aggregation and anti-thrombotic effects of Clove extracts were studied ex vivo methods by mesuring the inhibitory effect on thrombin induced platelet aggregation and the fibrinolytic activity. The Clove extracts were found to have a potent scavenging activity, as well as an inhibitory effect on LDL oxidation in vitro. Moreover Clove extracts were exhibited remarkable inhibitory effect on platelet aggregation and fibrinolytic activity. In conclusion, the Clove extracts have anti-oxidative and anti-atherosclerotic effects in vitro and ex vivo system, which can be used for developing pharmaceutical drug against oxidative stress and atherosclerosis.

• Journal of Oriental Neuropsychiatry
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• v.23 no.1
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• pp.115-128
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• 2012

Objectives : To evaluate the in vitro scavenging activity, inhibitory effect of LDL oxidation of pro-oxidant reactive species, anti-platelet aggregative effects and anti-thrombosis effects in response to treatment with SA using various screening methods including biological and non-biological oxidants. Methods : The antioxidant activity concerning extract from SA was studied with in vitro methods by measuring the antioxidant activity by TEAC, measuring the scavenging effects on reactive oxygen species (ROS) [superoxide anion, hydroxyl radical] and on reactive nitrogen species (RNS) [nitric oxide and peroxynitrite] as well as measuring the inhibitory effect on $Cu^{2+}$-induced human LDL oxidation and the inhibitory effect on thrombin-induced platelet aggregation and thrombosis. Results : SA extracts were found to have a potent scavenging activity regarding oxidative stress as well as an inhibitory effect towards LDL oxidation, platelet aggregation, and thrombosis. Conclusions : The SA extracts have anti-oxidative and anti-atherosclerotic effects in vitro system, which can be used for developing pharmaceutical drugs against oxidative stress and atherosclerosis.

• Microbiology and Biotechnology Letters
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• v.42 no.3
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• pp.302-306
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• 2014

Ehwa nuruk (EN), a traditional Korean alcoholic rice beverage, is manufactured from pulverized wet rice and the needles of pine trees. In this study, the ethanol extract of EN and its subsequent organic solvent fractions were prepared, and their in-vitro anti-thrombosis activity evaluated. In an anti-coagulation assay, only the ethylacetate (EA) fraction of the ethanol extract showed significant extensions of thrombin time, prothrombin time and activated partial thromboplastin time. In a platelet aggregation assay, the water residue of the ethanol extract exhibited aggregation inhibitory activity. Our results suggest that the EA fraction has potential as a new anticoagulation agent and EN could be used as a novel resource for anti-thrombosis agents. This report provides the first evidence of the anti-thrombosis activity of EN.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.5
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• pp.1494-1504
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• 2004

The present paper reports the effects of Hwaotang an atherosclerosis using a spontaneous experimental model, We have also investigated the pharmacological effect of Hwaotang on collagen- and ADP-induced blood platelet aggregation, thrombin-induced conversion of fibrinogen and fibrinolysis in in vitro experiments, and various effects on stimuli-induced superoxide generation in human neutrophils. Hwao-tang was shown to have inhibitory effect on collagen- and ADP-induced blood platelet aggregation, on thrombin-induced conversion of fibrinogen to fibrin and on the activity of plasminogen or plasmin. Hwao-tang also significantly inhibited fMLP-induced superoxide generation in a concentration-dependent manner, but not that induced by arachidonic acid. Hwao-tang inhibited neutrophil functions, including degranulation, superoxide generation, and leukotriene B4 production, without any effect on 5-lipoxygenase activity. In conclusion, the protection of extracts of Hwao-tang on the ischemic infarction induced artificially might be involved to their inhibition of thrombotic action. The results also indicate that Hwao-tang exerts the effects on superoxide generation related to the inhibition of neutrophil functions.

• Journal of Ginseng Research
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• v.45 no.4
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• pp.490-497
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• 2021

Background and objective: Synthetic ginsenoside compounds G-Rp (1,3, and 4) and natural ginsenosides in Panax ginseng 20(S)-Rg3, Rg6, F4 and Ro have inhibitory actions on human platelets. However, the inhibitory mechanism of ginsenoside Rk1 (G-Rk1) is still unclear thus, we initiated investigation of the anti-platelet mechanism by G-Rk1 from Panax ginseng. Methodology: Our study focused to investigate the action of G-Rk1 on agonist-stimulated human platelet aggregation, inhibition of platelet signaling molecules such as fibrinogen binding with integrin α_IIbβ₃ using flow cytometry, intracellular calcium mobilization, fibronectin adhesion, dense granule secretion, and thromboxane B2 secretion. Thrombin-induced clot retraction was also observed in human platelets. Key Results: Collagen, thrombin, and U46619-stimulated human platelet aggregation were dose-dependently inhibited by G-Rk1, while it demonstrated a more effective suppression on collagen-stimulated platelet aggregation using human platelets. Moreover, G-Rk1 suppressed collagen-induced elevation of Ca²⁺ release from endoplasmic reticulum, granule release, and α_IIbβ₃ activity without any cytotoxicity. Conclusions and implications: These results indicate that G-Rk1 possess strong anti-platelet effect, proposing a new drug candidate for treatment and prevention of platelet-mediated thrombosis in cardiovascular disease.

• Journal of Life Science
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• v.23 no.12
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• pp.1460-1470
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• 2013

To examine whether miscellaneous cereal grains have an antithrombotic effect, we investigated the anticoagulant activity of 80% ethanol extracts from eleven selected miscellaneous cereal grains. The 80% ethanol extract of hwanggeumchal sorghum (Sorghum bicolor) showed the highest anticoagulant activity, followed by that of green foxtail millet grains, in terms of thrombin time (TT). When the ethanol extract of hwanggeumchal sorghum was sequentially fractionated with n-hexane, methylene chloride, ethyl acetate, and n-butanol, the majority of the TT-inhibitory activity was detected in the hexane and methylene chloride fractions. Whereas aspirin (final conc. 480 ${\mu}g/ml$) prolonged TT by 2-fold, the ethanol extract, hexane fraction, and methylene chloride fraction in the same dose prolonged TT by 2.2-fold, 2.9-fold, and 2.5-fold, respectively. The ethanol extract of hwanggeumchal sorghum could delay activated partial thromboplastin time (APTT) as well as prothrombin time (PT). Although the APTT-inhibitory activity of the ethanol extract was mainly partitioned into the hexane and methylene chloride fractions, the PT-inhibitory activity of the ethanol extract was solely partitioned into the hexane fraction. The APTT- and PT-inhibitory activities of these organic solvent fractions were more potent than those of the control warfarin (final conc. 3.13 mg/ml). The TT-inhibitory activity of the ethanol extract was heat-stable and acid-stable. The ethanol extract, hexane fraction, and methylene chloride fraction of hwanggeumchal sorghum appeared to possess a direct fibrinolytic activity toward fibrin clotting. These results show that hwanggeumchal sorghum can exert anticoagulant and fibrinolytic effects and, thus, have the potential to be applicable as antithrombotic dietary sources.

• Korean Journal of Plant Resources
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• v.33 no.4
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• pp.237-244
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• 2020

The objective of this study was to identify and characterize fibrinolytic compound from Cornus officinalis. Cornus officinalis. Hot water extract was fractionated into hexane, chloroform, ethyl acetate, butanol, and water fractions. Assays for fibrinolytic activity indicated that only the ethyl acetate fraction had significant efficacy at 1.36 plasmin units/mL. Isolation of fibrinolytic compound was carried out on Amberlite IRA-400, Sephadex LH-20 and active charcoal column chromatography. HPLC analysis of the purified fibrinolytic compound showed retention time (RT) same as authentic malic acid. LC / MS / MS in negative mode showed the same peak at m/z 133, confirming that the purified compound was malic acid with a molecular weight 134 Da. The compound showed fibrinolytic activity of 0.69 plasmin units/mL, 14.62% of thrombin inhibitory activity, 6.42% of antioxidative activity, and 17.28% of α-glucosidase inhibitory activity. The purified compound hydrolyzed γ subunits of human fibrinogen. In conclusion, malic acid isolated from Cornus officinalis might have potential to be developed as ingredient for biofunctional foods to prevent cardiovascular diseases.

• Food Science and Preservation
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• v.17 no.4
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• pp.541-546
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• 2010

Wild-garlic (Allium victorialis L.) is a perennial plant found in worldwide and has been considered as a favorite vegetable due to its particular smell and taste. However, the study of biological activity of wild-garlic and the development of processed food are in rudimentary. In this study, we evaluated several biological activities, including antioxidant, antimicrobial, and inhibitory activities against human thrombin, $\alpha$-amylase and $\alpha$-glucosidase, of Ulrung wild-garlic. Analysis of the composition showed that Ulrung wild-garlic is nutritive although it is perishable. The color of fresh juice was stably maintained during 10 days-storage at $4^{\circ}C$, but rapidly discolored by heat treatment at $70^{\circ}C$ for 1 h. During heat treatment, the contents of total sugar and total polyphenol were decreased to 75% and 50%, respectively, and acidity was increased from 0.069% to 0.111%. In a while, the brix, reducing sugar, and total flavonoids showed minor changes. The fresh juice showed strong DPPH scavenging activity, reducing power and antibacterial and antifungal activity, but the heat-treated juice lost the antioxidant and antimicrobial activities. The inhibitory activities against human thrombin and $\alpha$-amylase and $\alpha$-glucosidase was negligible in both fresh juice and heat-treated juice. These results suggested that the antioxidant and antimicrobial components in wild-garlic are heat-liable and volatile. Based on our results, we propose non-heat treatment products for processed wild-garlic, for example, fresh juice-added beverage or fermented liquors using wild-garlic.

• Applied Biological Chemistry
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• v.42 no.4
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• pp.336-343
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• 1999

Fifty six mycelial cultured Basidiomycetes were screened for their inhibitory effects against prolyl endopeptidase(PEP), acetylcholine esterase(AChE) and thrombus coagulation. Out of them, methanolic extract of mycelium and/or ethylacetate(EtOAc) soluble fraction from culture broth of Peniophora quercina, Amanita aspera, Phellinus chrysoloma, Grifola frondosa, Wolfiporia extensa, Clavicorona pyxidata and Phanerochaete sordida inhibited more than 90% of PEP activity at 40 ppm. The extracts of Lenzites betulina, Phellinus chrysoloma, Wolfiporia extensa, Phanerochaete sorrlida, Hypocrea nigricans, Coriolus azureus, Flammulina velutipes, Phlebiopsis gigantea and Bondarzewia montana exhibited about 40% of inhibitory activity against AChE at 40 ppm. In thrombin times assay, the extracts of Amanita aspera, Oxyporus latemarginata, Peniophora quercina, Fomes fomenfarius, Trametes versicolor, and Phlebiopsis gigantea delayed coagulation of thrombus about two to three times over control at ca 550 ppm. In activated partial thromboplastin times assay, none of the tested Basidiomycetes showed significant effect.