CAJ | 학술논문

背景:蝰亚科、蝮亚科和眼镜蛇科等蛇毒中存在着一类能直接溶解纤维蛋白(原)的酶,称为纤维蛋白(原)溶酶(纤溶酶),可做为溶解在心肌梗死、中风等期间形成的血栓的抗栓药物.目的:分离纯化江浙蝮蛇毒纤溶酶,并分析其部分生物化学和药理性质.设计:对照观察实验.单位:广西医科大学蛇毒研究所.材料:实验于2003-07/2005-06在广西医科大学蛇毒研究所完成.昆明种小鼠,体质量18～25 g,平均(20±2)g,鼠龄三四个月,由广西医科大学实验动物中心提供.蛇毒冻干粉由广西医科大学蛇毒研究所提供.方法:应用DEAE Sepharose CL-6B阴离子交换柱和HiPrep Sephacryl S100柱分离纯化纤溶酶,以高效液相色谱仪和聚丙烯酰胺凝胶电泳检测纯度、相对分子质量,应用等电聚焦电泳测定等电点,然后测定氨基酸组分及部分药理性质.主要观察指标:纤溶酶相对分子质量、等电点和氨基酸组成,抑制剂对纤溶酶的作用,出血活性实验,纤溶酶对家兔体内抗凝活性的作用.结果:纳入30只鼠和12只兔全部进入结果分析.分离得到相对分子质量为59 100,等电点为4.98,SDS-PAGE为一条带,高效液相色谱分析为单一峰的纤溶酶.该组分属于金属蛋白酶类.氨基酸组成分析表明它含较多的酸性氨基酸.有微弱的出血毒性.具有较强且持久的溶栓效果.结论:分离纯化得到了一种有微弱出血毒,具有较强抗凝活性的单一组分的酸性金属蛋白酶.
배경:호아과、복아과화안경사과등사독중존재착일류능직접용해섬유단백(원)적매,칭위섬유단백(원)용매(섬용매),가주위용해재심기경사、중풍등기간형성적혈전적항전약물.목적:분리순화강절복사독섬용매,병분석기부분생물화학화약이성질.설계:대조관찰실험.단위:엄서의과대학사독연구소.재료:실험우2003-07/2005-06재엄서의과대학사독연구소완성.곤명충소서,체질량18～25 g,평균(20±2)g,서령삼사개월,유엄서의과대학실험동물중심제공.사독동간분유엄서의과대학사독연구소제공.방법:응용DEAE Sepharose CL-6B음리자교환주화HiPrep Sephacryl S100주분리순화섬용매,이고효액상색보의화취병희선알응효전영검측순도、상대분자질량,응용등전취초전영측정등전점,연후측정안기산조분급부분약이성질.주요관찰지표:섬용매상대분자질량、등전점화안기산조성,억제제대섬용매적작용,출혈활성실험,섬용매대가토체내항응활성적작용.결과:납입30지서화12지토전부진입결과분석.분리득도상대분자질량위59 100,등전점위4.98,SDS-PAGE위일조대,고효액상색보분석위단일봉적섬용매.해조분속우금속단백매류.안기산조성분석표명타함교다적산성안기산.유미약적출혈독성.구유교강차지구적용전효과.결론:분리순화득도료일충유미약출혈독,구유교강항응활성적단일조분적산성금속단백매.
BACKGROUND: A fibrinolytic enzyme (FLE) was purified from the venom of Agkistrodon blomhoffi brevicaudus by ion-exchange chromatography;meanwhile, it can dissolve thrombotic anti-thrombus drugs during onset of myocardial infarction and apoplexy.OBJECTIVE: To investigate the purification of FLE from the venom of Agkistrodon blomhoffi brevicaudus and research some biochemical and pharmacological characterizations.DESIGN: Controlled observational study.SETTING: Snake Venom Research Institute, Guangxi Medical University.MATERIALS: The experiment was carried out in Snake Venom Research Institute of Guangxi Medical University from July 2003 to June 2005.Kunming mice, weighing 18-25 g, of mean body mass of (20±2) g, aged 3-4 months were provided by Experimental Animal Center of Guangxi Medical University. Agkistrodon blomhoffi brevicaudus was supplied by Snake Venom Research Institute of Guangxi Medical University.METHODS: FLE was purified with DEAE Sepharose CL-6B ion exchang columns and HiPrep Sephacryl S 100; purity and relative molecular mass were measured with high-effective chromatograph of liquid and polyacrylamidedel electrophoresis (PAGE); isoelectric point was measured with isoelectric focusing electrophoresis; and then, components and partially pharmacological characteristics were also measured.MAIN OUTCOME MEASURES: Relative molecular mass, conformation of isoelectric point and amino acid, effect of inhibitor on FLE, hemorrhagic activity, and anticoagulant effect of FLE.RESULTS: All 30 rats and 12 rabbits were involved in the final analysis.Relative molecular mass was 59 100 and isoelectric point was 4.98. SDSPAGE acted as a zone in vitro. High-effective chromatograph of liquid showed a simple peak of FLE. This group belonged to metallo proteinases.Analysis of components of amino acid suggested that it contained a lot of acidity amino acids, had mild hemorrhagic toxicity and strongly thrombolytic effect.CONCLUSION: A simple acidity metallo proteinases, which is characterized by mild hemorrhagic toxicity and strong anti-coagulation, can be purified from the venom of Agkistrodon blomhoffi brevicaudus.