• Title/Summary/Keyword: Fibrin plate

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

  • 김준호
    • 대한의생명과학회지
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    • 제13권3호
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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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뽕나무버섯으로부터 Fibrinolytic enzyme의 정제 및 특성 연구 (Purification and Characterization of Fibrinolytic Enzyme from Armillariella mellea)

  • 김준호;김양선
    • 한국균학회지
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    • 제26권4호통권87호
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    • pp.583-588
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    • 1998
  • 뽕나무버섯 추출물에서 fibrinolytic enzyme을 분리 정제하였다. 이 효소의 분자량은 약 19800Da 이고 pH 7.0에서 최고의 활성을 보여주는 neutral protease로 $55^{\circ}C$에서 최대 활성을 나타냈다. EDTA에 의해 활성이 저해되는 것으로 보아 metalloprotease로 추정되며 $Hg^{+2}$의 영향을 받아 활성이 상실되었다. N-terminal amino acid 분석 결과 15번째까지의 아미노산 잔기 순서는 X-X-T-N-G-X-T-X-S-R-Q-T-T-L-V 이었으며 지금까지 알려진 것과는 다른 새로운 효소이다.

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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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    • 제8권2호
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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.

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

  • 정순모;송효남
    • 동아시아식생활학회지
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    • 제19권3호
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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
    • 대한약침학회지
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    • 제14권3호
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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.

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

  • 조은경;정유정;갈상완;최영주
    • 생명과학회지
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    • 제19권10호
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    • pp.1424-1431
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    • 2009
  • 전통발효식품인 새우젓으로부터 혈전과 chitin 분해력이 우수한 균을 분리하였으며 16S rRNA 염기서열 분석으로 B. licheniformis와 가장 유사한 균주임을 확인하였다. 이 균주를 B. licheniformis SC082로 명명하였고, 최적 생육조건은 $37^{\circ}C$, pH 7.0, 염 농도 6%로 확인되었다. 이 균주의 기질특이성을 조사한 결과, 우수한 혈전분해력을 나타냈으며 1% 농도의 colloidal chitin의 첨가에 의하여 chitinase 활성이 증가됨을 관찰할 수 있었다. 또한 지질 분해능은 없었고 약한 skim milk 분해력을 가지고 있었다. SDS-PAGE와 zymogram 분석 결과, 이 균은 혈전분해효소 isozyme과 chitinase isozyme을 생성하는 균주로 확인되었다. 그 대략적인 분자량은 각각 22.0, 66.0, 72.0 kDa과 55.0, 62.0 kDa이었다. B. licheniformis SC082 균주가 생성하는 혈전분해효소는 pH 9.0 그리고 $50^{\circ}C$까지 안정하게 유지되었고, 이와 더불어, chitinase 활성은 pH 5, $45^{\circ}C$일 때 높게 나타났다. B. licheniformis SC082 균주의 DPPH 전자공여능법에 의한 항산화력은 농도의 증가에 따라 상승되었는데 $20\;{\mu}g$의 균상등액에 대한 항산화력은 약 31%으로 나타났다.

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

  • 김은정;김준호
    • 한국버섯학회지
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    • 제13권1호
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    • pp.30-36
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    • 2015
  • 아위버섯을 혈관계질환 예방과 치료를 위한 기능성 식품 개발에 이용하기 위한 기초 자료를 얻기 위해 아위버섯 물추출물과 유기용매 분획물의 혈전용해활성과 트롬빈 저해활성, 항산화활성 및 항염증활성을 확인하였다. 에틸아세테이트 분획물은 1.33 plasmin unit의 높은 혈전용해 활성과, 94.45%의 높은 트롬빈 저해활성을 나타냈으며, 물 추출물은 37.01%의 항산화활성을 나타냈으며, 클로로포름 분획물은 98.13%의 가장 높은 항염증활성을 나타냈다. 실험 결과로부터 아위버섯의 에틸아세테이트 분획물은 높은 혈전용해활성, 트롬빈 저해활성 그리고 클로로포름 분획물은 높은 항염증활성을 나타내 심혈관계 질환 관련 치료나 예방을 위한 기능성식품 개발에 이용 가능할 것으로 기대된다.

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

  • 김준호;조승구
    • 한국균학회지
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    • 제32권2호
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    • pp.119-124
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    • 2004
  • 쓴송이버섯으로부터 분리한 혈전용해효소(TSFE)의 활성은 11.42 U/mg이었으며, N-terminal amino acid 서열은 Ala-Thr-Tyr-Lys-Ile-X-Ser-Ala-Thr-His-Gln-X-X-Leu-Val로 지금까지 발표되지 않은 새로운 효소였다. MALDI-TOF와 ICP/MS로 분자량은 18788.25 Da이며, $Zn^{2+}$을 함유하는 금속효소임을 알게 되었다. 이 효소는 $40^{\circ}C$까지 열에 안정하고, 특히 합성된 기질 Lys pNA를 강하게 분해하였다. $Zn^{2+}$$Co^{2+}$에 의해 활성이 증가되고, EDTA, 1,10-phenanthroline, $Hg^{2+}$에 의해서는 활성이 완전히 소멸되었다. 이 효소는 섬유소원의 $A{\alpha}$ chain은 분해하지만, $B{\beta}$ chain과 ${\gamma}$ chain은 분해하지 못했다.

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

  • 양원경;성윤영;전명숙;윤태숙;김호경
    • 대한본초학회지
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    • 제25권4호
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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.

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

  • 김현애;이상한;최영철;박관호;황재삼;김남정;남성희
    • 한국잠사곤충학회지
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    • 제51권2호
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    • pp.147-152
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    • 2013
  • 국내 유용 곤충의 70% 에탄올 및 물 추출물의 혈전용해 효과를 확인하기 위하여 fibrin plate를 이용하여 혈전용해능을 측정하였고, 그 분해산물을 SDS-PAGE를 이용하여 분리하였다. 그 결과 5종 (방울벌레 유충, 방울벌레 성충, 장수풍뎅이 성충, 꽃무지 유충, 꿀벌 유충)의 물 추출물에서 plasmin과 비교하여, 높은 혈전 용해 능을 나타내었고, 분해 패턴 또한 plasmin과 비슷하거나 더 강한 단백질 분해능을 나타내어 물 추출물에서 높은 혈전 용해능을 확인할 수 있었다.