关键词: 眼镜蛇毒，蛋白酶，纤维蛋白原，血小板聚集

摘要：目的：从中国眼镜蛇毒中分离纯化出可水解纤维蛋白原的蛇毒蛋白酶，并观察其体内给药对血小板聚集的影响。方法：使用超滤的方法以及肝素亲和层析柱HeparinSepharoseCL-6B和凝胶层析柱SephadexG150分离纯化中国眼镜蛇毒蛋白酶。SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)检测其纯度和分子量。比浊法测定血小板聚集率。结果：从中国眼镜蛇毒中纯化出具有水解纤维蛋白原活性的蛋白酶，经SDS-PAGE电泳测定为一条带，分子量为47.1kD。该蛋白酶低剂量(0.025mg／kg)、中剂量(0.05mg／kg)以及高剂量(0.1mg／kg)体内给药后，剂量依赖性地抑制ADP(10μmol／L)、胶原(100μg／ml)诱导的兔血小板聚集。结论：具有水解纤维蛋白原活性的中国眼镜蛇毒蛋白酶在体内表现出抑制血小板聚集的作用。

分类号：R973.2

Isolation and Purification of Chinese Cobra Venom Proteinase and Its Effect on Platelet Aggregation

guo Dan,Wu Shuguang,Zhu Zhengguang,Chen Nana(Pharmaceutical Department of Nan fang Hospital, NO.1 Military Medical University Guangzhou 510515)

Abstract：Purpose: To isolate and purify the proteinase which degrades fibrinogen from Chinese cobra venom (Naja naja atra) and investigate its effects on the platelet aggregation in vivo. Methods: Isolation and purification of Chinese cobra venom proteinase were carried out by ultrafiltration and successive chromatography on Heparin Sepharose CL-6B column and Sephadex Gl50 column. Homogenicity and molecular weight of the proteinase were determined in SDSpolyacrylamide gel electrophoresis (SDS-PAGE). Platelet aggregation was evaluated. Results: Chinese cobra venom proteinase which degrades fibrinogen was a single peptide-chain with a molecular mass of about 47 100 dalton as revealed by SDS-PAGE. It dose-dependently inhibited rabbit platelet aggregation induced by 10 μmol/L of ADP or 100μg/ml of collagen when tested at the dosages of 0.025, 0.05 and 0.1mg/kg. Conclusion: Chinese cobra venom proteinase which degrades fibrinogen exerted an inhibitory effect on platelet aggregation in vivo.

Keywords：Cobra venom, Proteinase, Fibrinogen, Platelet aggregation

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收稿日期：1999-09-20

修稿日期：1999-11-29