• Title/Summary/Keyword: Fibrin plate

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Screening Test of Wild Mushroom Methanol Extracts for Fibrinolytic and $\alpha-Glucosidase$ Inhibitory Activity

  • Kim, Jun-Ho;Yoo, Kwan-Hee;Seok, Soon-Ja
    • Biomedical Science Letters
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    • v.13 no.3
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    • pp.245-249
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    • 2007
  • We investigated the fibrinolytic and ${\alpha}-glucosidase$ inhibitory activity of 55 wild mushroom methanol extracts. Among them, 14 mushrooms showed fibrinolytic activity. In particular, Amanita virgineoides showed the greatest enzyme activity (3.9 plasmin units/ml) by a fibrin plate assay. The fibrinolytic activities of Suillus pictus and Polypolellus varius were 3.8 plasmin units, and the activity of Gomphus fujisanensis was 2.8 plasmin units. Leccinum extremiorientale and Xerocomus nigromaculatus had the same activity with 2.3 plasmin units. In a ${\alpha}-glucosidase$ inhibitory activity test, Lactarius sp. showed the greatest inhibitory activity at 97.3%. The ${\alpha}-glucosidase$ inhibitory activities of Clitocybe odora, Xerocomus nigromaculatus, Melanoleuca melaleuca, Suillus pictus, and Gyroporus castaneus were 84.3%, 77.9%, 74.6%, 68.7%, and 65.4%, respectively. According to the results, because Suillus pictus and Xerocomus nigromaculatus have strong fibrinolytic and ${\alpha}-glucosidase$ inhibitory activities, the two mushrooms will be used as materials for the development of new biofunctional food.

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Purification and Characterization of Fibrinolytic Enzyme from Armillariella mellea (뽕나무버섯으로부터 Fibrinolytic enzyme의 정제 및 특성 연구)

  • Kim, Jun-Ho;Kim, Yang-Sun
    • The Korean Journal of Mycology
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    • v.26 no.4 s.87
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    • pp.583-588
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    • 1998
  • A fibrinolytic enzyme has been isolated from the edible honey mushroom, Armillariella mellea and purified. The apparent molecular mass of purified enzyme was estimated to be 19800Da by SDS polyacryl amide gel electrophoresis and 19900Da by gel filtration, indicating that it was a monomer. The enzyme was optimal at pH 7, suggesting that the purified enzyme was a neutral proteinase. It shows the maximum fibrinolytic activity at $55^{\circ}C$, is completely inactivated above $65^{\circ}C$, and still indicates 40% of activity at $37^{\circ}C$. The fibrinolytic activity has been decreased by the addition of EDTA. Fifteen amino acid sequence was determined by protein sequencing techniques.

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Effects of Insect Crude Drugs on Blood Coagulation and Fibrinolysis System

  • Ahn, Mi-Young;Hahn, Bum-Soo;Ryu, Kang-Sun;Cho, Sung-Ig
    • Natural Product Sciences
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    • v.8 no.2
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    • pp.66-70
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    • 2002
  • The in vitro anticoagulant and fibrinolytic activities of crude extracts from insects were evaluated in order to find effective therapeutic drugs for the treatment of myocardial and cerebral thrombosis. We prepared three types of extracts (water, methanol and ethylacetate) from 28 insects for use as raw materials for the activity assays. The fibrinolytic activity was tested using the fibrin plate method and the activated partial thromboplastin time and thrombin time were measured for blood clotting activity. With regards to the fibrinolytic system, water extracts of six kinds of insects displayed a remarkable level of activity with a plasmin-like action. The water extracts of [Catharsius molossus, Eupolyphaga sinensis, Huechys sanguinea, Mantidis $o\ddot{o}theca$, Mimela splendens, and Polistes mandarinus (Vespae Nidus)] exhibited the activity. On the other hand, the methanol extracts did not display any fibrinolytic activity. In terms of the coagulation system, an aqueous extract of silkworm Tongchunghacho (Paecilomyces japonica), Oxya japonica japonica and Buthus martensi (Scorpion) increased the clotting time significantly longer (181 times) than the control. These results suggest that crude drugs from insects are useful sources for the development of new drugs for use in treatments involving blood coagulation and fibrinolysis.

Biological Activities of Fermented Mugworts and Their Effects on Lipid Metabolism in Rats (발효쑥의 생리활성 및 흰쥐의 지질대사에 미치는 영향)

  • Jung, Soon-Mo;Song, Hyo-Nam
    • Journal of the East Asian Society of Dietary Life
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    • v.19 no.3
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    • pp.356-362
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    • 2009
  • The objective of this study was to investigate the biological activities of fermented mugworts and their effects on the lipid metabolism in hyperlipidemia-induced rats. The proximate compositions of two kinds of mugwort, Artemisia capillaris Thumberg and Artemisiae asiaticae Nakai, were compared before and after fermentation. In both types of mugwort crude protein and amino nitrogen contents markedly increased with fermentation. Thrombolytic activity determined from the size of the clear zone on a fibrin plate was higher for the Artemisiae asiaticae Nakai, especially in the fermented sample. Antioxidative activity according to DPPH(1,1-diphenyl-2-picrylhydrazyl) radical scavenging effects was higher for Artemisia capillaris Thumberg but no significant differences were found between the samples after fermentation. The body weights of hyperlipidemia-induced rats that were fed the mugworts for 4 weeks were lower than those of the control group. In all the mugwort-fed rats serum total cholesterol and LDL-cholesterol levels remarkably decreased. Furthermore the fermented mugworts were found to be more effective at decreasing triglyceride (TG) levels. It is also noteworthy that the highest HDL-cholesterol levels were observed in the rats treated with the fermented Artemisiae asiaticae Nakai. And fatty liver weights were higher in the rats fed Artemisia capillaris Thumberg. In conclusion the feeding of fermented Artemisiae asiaticae Nakai was effective for increasing thrombolytic activity and HDL-cholesterol levels as well as reducing TG levels in rats.

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Characterization of Fibrinolytic Proteases from Gloydius blomhoffii siniticus Venom

  • Choi, Suk-Ho
    • Journal of Pharmacopuncture
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    • v.14 no.3
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    • pp.71-79
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    • 2011
  • Objectives : This study was undertaken to identify fibrinolytic proteases from Gloydius blomhoffii siniticus venom and to characterize a major fibrinolytic protease purified from the venom. Methods : The venom was subjected to chromatography using columns of Q-Sepharose and Sephadex G-75. The molecular weights of fibrinolytic proteases showing fibrinolytic zone in fibrin plate assay were determined in SDS-PAGE (Sodium dodecyl sulfate-polyacrylamide gel electrophoresis) The effects of inhibitors and metal ions on fibrinolytic protease and the proteolysis patterns of fibrinogen, gelatin, and bovine serum albumin were investigated. Results : 1) The fibrinolytic fractions of the three peaks isolated from Gloydius blomhoffii siniticus venom contained two polypeptides of 46 and 59 kDa and three polypeptides of 32, 18, and 15 kDa and a major polypeptide of 54 kDa, respectively. 2) The fibrinolytic activity of the purified protease of 54 kDA was inhibited by metal chelators, such as EDTA, EGTA, and 1,10-phenanthroline, and disulfhydryl-reducing compounds, such as dithiothreitol and cysteine. 3) Calcium chloride promoted the fibrinolytic activity of the protease, but mercuric chloride and cobalt(II) chloride inhibited it. 4) The fibrinolytic protease cleaved preferentially A${\alpha}$-chain and slowly B${\beta}$-chain of fibrinogen. It also hydrolyzed gelatin but not bovine serum albumin. Conclusions : The Gloydius blomhoffii siniticus venom contained more than three fibrinolytic proteases. The major fibrinolytic protease was a metalloprotease which hydrolyzed both fibrinogen and gelatin, but not bovine serum albumin.

Isolation and Characterization of Bacillus licheniformis SC082 Degrading Fibrin and Chitin from Shrimp Jeot-Gal (새우젓으로부터 혈전과 chitin 분해능을 지닌 균주 Bacillus licheniformis SC082의 분리 및 특성)

  • Cho, Eun-Kyung;Jung, Yu-Jung;Gal, Sang-Wan;Choi, Young-Ju
    • Journal of Life Science
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    • v.19 no.10
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    • pp.1424-1431
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    • 2009
  • Shrimp Jeot-Gal is a popular traditional Korean fermented seafood and has been used for seasoning. We isolated a bacterium showing strong extra-cellular fibrinolysis and chitinase activity from shrimp Jeot-Gal and the strain was designated SC082. SC082 was identified as Bacillus licheniformis by 16S rRNA sequence homology search. B. licheniformis SC082 exhibited optimum temperature, pH, and salt concentration at $37^{\circ}C$, pH 7.0, and 6%, respectively. Substrate specificity of the culture supernatant from B. licheniformis SC082 was detected in fibrin, skim milk, and chitin plate. The fibrinolytic activity was highly maintained up to $50^{\circ}C$ at a pH of 7.0 for 3 hr and was stable up to pH 9.0 at $37^{\circ}C$ for 3 hr. The chitinase activity was remarkably induced by addition of 1.0% colloidal chitin and the pH and temperature optima of the enzyme were 5.0 and $45^{\circ}C$, respectively. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis and zymogram analysis, this strain produced three fibrinolytic isozymes and two chitinase isozymes. The approximate molecular weights of the putative fibrinolytic enzymes were 23.0, 62.0, and 72.0 kDa and those of the chitinases were 62.0 and 55.0 kDa, respectively. The antioxidant activity of SC082 was also measured by using 2,2-diphenyl-l-picryl-hydrazyl (DPPH) free radical. The DPPH radical scavenging was slightly increased in a dose-dependent manner.

Fibrinolytic, thrombin inhibitory, anti-oxidative and anti-inflammatory activities of Pleurotus ferulea (아위버섯 추출물의 혈전용해, 트롬빈저해, 항산화 및 항염증 활성)

  • Kim, Eun-Jung;Kim, Jun-Ho
    • Journal of Mushroom
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    • v.13 no.1
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    • pp.30-36
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    • 2015
  • Our study investigated the fibrinolytic, thrombin inhibitory, anti-oxidative and anti-inflammatory activities of the water extract and solvent fractions isolated from Pleurotus ferulea. Fibrinolytic activity was investigated using the fibrin plate method. Thrombin inhibitory activity was used to analyze thrombin inhibitor assay. The DPPH assay was used to estimate anti-oxidative activity. Inhibition of NO production was measured for anti-inflammatory activity in LPS-activated murine RAW 264.7 macrophage cells. An MTS assay was used to evaluate the effects of the water extract and solvent fractions isolated from Pleurotus ferulea on cell viability. Our results showed the fibrinolytic activity to be strong in the ethyl acetate fraction at 1.33 plasmin units. The ethyl acetate fraction also showed high thrombin inhibitory activity at 94.45%. The anti-oxidative activity of the water extract was 37.01% and the anti-inflammatory activity of the chloroform fraction was 98.13%. These findings suggest that Pleurotus ferulea's extract and fractions could be applicable in the development of functional foods for the treatment and prevention of cardiovascular diseases.

Characterization of a Fibrinolytic Metalloenzyme from a Wild Mushroom, Tricholoma sejunctum (쓴송이버섯으로부터 분리한 혈전용해 금속효소의 특성 연구)

  • Kim, Jun-Ho;Cho, Seung-Koo
    • The Korean Journal of Mycology
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    • v.32 no.2
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    • pp.119-124
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    • 2004
  • Metalloenzyme was purified from the fruiting bodies of Tricholoma sejunctum. MALDI-TOF and ICP/MS analyses revealed that the enzyme had a molecular weight of 18788.25 and includes $Zn^{2+}$ ion. The N-terminal amino acid sequence of the enzyme was Ala-Thr-Tyr-Lys-Ile-X-Ser-Ala-Thr-His-Gln-X-X-Leu-Val. The activity of the enzyme was inhibited by EDTA and 1,10-phenanthroline, indicating that the enzyme was a metalloprotease. No inhibition was found with E-64 and pepstatin. It has broad substrate specificity for synthetic peptides. The enzyme was stable up to $40^{\circ}C$. The activity of the enzyme was increased by $Zn^{2+}$ and $Co^{2+}$, while it was totally inhibited by $Hg^{2+}$. The enzyme hydrolyzes $A{\alpha}$ subunit of human fibrinogen but did not show any reactivity for $B{\beta}$ and ${\gamma}$ form of human fibrinogen.

Comparative Study of Extracts from Rhubarb on Anti-Thrombotic and Anti-Platelet Activity (4종 대황(大黃)류 추출물의 항혈전 효능 비교)

  • Yang, Won-Kyung;Sung, Yoon-Young;Cheon, Myeong-Sook;Yoon, Tae-Sook;Kim, Ho-Kyoung
    • The Korea Journal of Herbology
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    • v.25 no.4
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    • pp.1-6
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    • 2010
  • Objective : The purpose of this study is to compare anti-thrombotic activities of 4 Rhubarb species. Methods : Rhubarb has been used as medical and edible resources worldwide for the therapy of cancer, constipation and inflammation, etc. To compare the anti-thrombotic activities among the species within rhubarb, we investigated and compared the inhibitory properties of water extracts from R. palmatum(RPE), R. officinale(ROE), R. nobile(RNE), and R. franzenbachii (RFE) on the fibrinolysis, blood coagulation and platelet aggregation. APTT and PT, blood coagulation time, were measured by apparatus of auto blood coagulation analysis that called ACL-7000. The measurement of fibrinolysis was measured and compares with the width of clear zone from melted fibrin plate. The platelet aggregation was measured and compared with inhibition rate of aggregation. Results : We found out the best species from rhubarb showing the significant fibrinolytic, anti-coagulant and anti-platelet aggregation activity. Our results show that Rheum nobile (RNE) has the most effective anti-thrombotic activity among 4 tested rhubarb. Conclusions : This study, therefore, RNE will be able to be used the development of drug which is thrombosis treatment.

Comparison of fibrinolytic activity from Korean indigenous insects (국내 토착 곤충의 항혈전 비교)

  • Kim, Hyunae;Lee, Sang Han;Choi, Youngcheol;Park, Kwanho;Hwang, Jaesam;Kim, Namjung;Nam, Sunghee
    • Journal of Sericultural and Entomological Science
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    • v.51 no.2
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    • pp.147-152
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    • 2013
  • The fibrinolytic activity of aqueous extracts from Korean indigenous insects were studied. Fibrinolytic activity of aqueous extract from 5 insects (larva and adult parts of Meloimorpha japonica, Allomyrina dichotoma, Cetonia pilifera, Apis mellifera) showed 2.7-fold potent than that of plasmin used as a positive control. In addition, the fibrinolytic activity was observed through the degradation products by SDS-PAGE. The extracts efficiently hydrolyzed ${\alpha}-$, ${\beta}-$ and ${\gamma}$ chains of human fibrinogen. This study suggested that aqueous extracts from Korean indigenous insects have potential in developing a useful source of antithrombosis agent(s).