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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18～25g,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DEAESepharoseCL-6B阴离子交换柱和HiPrepSephacrylS100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59100,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。
BACKGROUND: There exists a class of enzymes that directly dissolve fibrin (prot), such as fibrin (procollagen) (plasmin), in the venoms of the subfamily Subfamily, Subfamily, and Cobra, Antithrombotic drugs that form thrombus during myocardial infarction, stroke, and the like. OBJECTIVE: To isolate and purify the fibrinolytic enzyme of viper venom from Jiangsu and Zhejiang and to analyze its biochemical and pharmacological properties. Design: Control observation experiment. Unit: Guangxi Medical University Institute of snake venom. MATERIALS: Experiments were performed at the Institute of Snake Venom, Guangxi Medical University from July 2003 to June 2005. Kunming mice, body weight 18 ~ 25g, an average of (20 ± 2) g, three or four months old, provided by the Experimental Animal Center of Guangxi Medical University. Snake venom freeze-dried powder provided by the Guangxi Medical University Institute of snake venom. Methods: The plasmin was separated and purified by DEAE Sepharose CL-6B anion exchange column and HiPrepSephacryl S100 column. The purity and relative molecular mass were determined by high performance liquid chromatography and polyacrylamide gel electrophoresis. The isoelectric point was determined by isoelectric focusing electrophoresis. Amino acid components and some pharmacological properties. MAIN OUTCOME MEASURES: The relative molecular mass of plasmin, isoelectric point and amino acid composition, the effect of inhibitor on plasmin, hemorrhagic activity, the effect of plasmin on anticoagulant activity in rabbits. RESULTS: Thirty rats and 12 rabbits were included in the result analysis. The relative molecular mass was 59100, isoelectric point was 4.98, SDS-PAGE as a band, high performance liquid chromatography analysis of a single peak of plasmin. This component belongs to the class of metalloproteases. Amino acid composition analysis showed that it contains more acidic amino acids. A slight bleeding toxicity. Has a strong and lasting thrombolytic effect. CONCLUSION: A single component acidic metalloprotease with a weak bleeding toxicity and strong anticoagulant activity was isolated and purified.