江浙蝮蛇毒抗血小板聚集组分的分离纯化及活性分析

蛇毒，特别是蝰蛇和眼镜蛇家族蛇毒含有相当数量作用于血小板的活性成分，它们可诱导或抑制血小板聚集。一些相关成分应用于临床有望作为有效药物治疗出血或血栓性疾病。江浙蝮蛇属蝰蛇类，是我国特有的蛇种。本研究以是否抑制ADP诱导的血小板聚集作为示踪，从江浙蝮蛇毒中分离纯化出一种血小板聚集抑制剂并对其生化特性做进一步探讨。江浙蝮蛇毒经DEAE-Sepharose CL-6B，Sephadex G-75及在FPLC上利用SP-Sepha-rose和Mono Q层析，得到SDS-PAGE条带均一、分子量为65kD的蛋白质组分。该物质无磷脂酶A2、精氨酸酯酶及纤溶酶活性，具有拮抗ADP、胶原诱导的血小板聚集作用。结论：分子量为65kD的蛋白质组分，可抑制ADP、胶原诱导的血小板聚集，且呈明显的剂量一效应关系。

Purification and Characterization of Platelet Aggregation Inhibitor Component from Venom of Agkistrodon Halys Pallas

Snake venom proteins,particularly from the viper and elapid families,have been known to contain a number of platelet active components including what cause platelet aggregation or inhibit platelet aggregation. Some of them have potential clinical usefulness for the treatment of human hemorrhagic or thrombotic disease. Agkistrodon halys pallas belonging to viper family is only growing in China. The aim of this study was to purify a human platelet aggregation inhibitor from venom of Agkistrodon halys pallas and determine its biochemical character. Whether a component could inhibit human platelet aggregation was act as a method to follow the tracks of the protein. Crude venom of Agkistrodon halys pallas was loaded onto a DEAE Sepharose CL 6B chromatography column could gain 6 peaks. A platelet inhibitor with molecular mass of 65 kD on SDS PAGE, was purified from peak 2 by Sephadex G 75 gel filtration and SP Sepharose, Mono Q on FPLC. It could inhibit human platelet aggregation induced by ADP, collagen without activities of phospholipase A2, esterase, fibrinogenolytic. It is concluded that a platelet inhibitor can be isolated and purified from venom of Agkistrodon halys pallas and its inhibition of platelet aggregation is does dependent.