广东眼镜王蛇毒酸性磷脂酶A_2的分离纯化及抗血小板聚集作用  

Isolation and Purification of Acid PLA_2 from Ophiophagus hannah Venom and Its Inhibition on Platelet Aggregation

作　　者：张维文[1] 钟倍华 张贵平 

机构地区：[1]广州医学院药理教研室,广州510182 [2]广州蛇毒研究所,广州510182

出　　处：《蛇志》2000年第1期3-6,共4页Journal of Snake

摘　　要：目的　研究眼镜王蛇毒中酸性磷脂酶 A2 对血小板的作用。方法　采用 CM-Sephadex C-2 5,Sephadex G-75,DEAE-Sephadex A-2 5,Sephadex G-75多步柱层析法 ;聚丙烯酰胺凝胶电泳 ;酸性磷脂酶 A2 酶活性测定 ;血小板聚集实验。结果　从粗毒中分离纯化出一酸性磷脂酶 A2 单体 ,分子量为 1 4KD;等电点 p I3 .8;PLA2 比活力 2 0 μmol· mg- 1 ·min- 1 ;小鼠腹腔注射 LD5 0 为 ( 7.53± 1 .2 9) mg.kg- 1 ;对 ADP、花生四稀酸、凝血酶诱导的兔血小板聚集均有抑制作用 ,具有剂量依赖关系。结论　广东眼镜王蛇毒酸性磷脂酶单体具有抗血小板聚集的作用。Objective This paper reports a potent platelet aggregation inhibitor which was an acidic PLA 2 purified from Ophiophagus hannah venom Methods Acidic PLA 2 was obtained from Guangdong Ophiophagus hannah venom by column chromatography and rechromatography on CM Sephadex C 25, Sephadex G 75, DEAE Sephadex A 25 The purity was tested by polyacrylamid gel electrophoresis ,gel isoelectric focusing electrophoresis and SDS electrophoresis; Molecular weight was determined by SDS PAGE The activity measurement of PLA 2 was carried out by automatic titration;The experiment of platelet aggregation was by the turbidimetric method  Results  Acidic PLA 2 was purified as a monomer, pI 3 8, MW 14KD, PLA 2 enzyme activity 20 μmol·mg 1 ·min 1 ,LD 50 (ip)7 53±1 29mg kg 1 Inhibition of rabbit platelet aggregation induced by ADP,arachidonic acid(AA),thrombin(thr) was dose dependent  Conclusion Acidic PLA 2 from Ophiophagus hannah venom could inhibit platelet aggregation induced by ADP,AA,thr

关 键 词：眼镜王蛇 蛇毒 酸性磷脂酶A2 血小板 聚集作用 

分 类 号：R996.3[医药卫生—毒理学] Q556.103[医药卫生—药学]