Hemorrhagic activity and mechanism of FIIa,a fibrinolytic enzyme from Agkistrodon acutus venom

作     者：Qing-qing WANG2, Jia-shu CHEN, Xiu-xia LIANG3, Peng-xin QIU, Yi-wen WANG, Guang-mei YAN Department of Pharmacology, Zhongshan Medical College, Sun Yat-Sen University, Guangzhou 510080, China Now in Beijing Universityof Chinese Medicine,Beijing 100029,China. 

作者机构：Department of Pharmacology Zhongshan Medical College Sun Yat-Sen University 

出 版 物：《Acta Pharmacologica Sinica》 (中国药理学报(英文版))

年 卷 期：2004年第25卷第4期

页      面：116-123页

核心收录：

学科分类：1007[医学-药学(可授医学、理学学位)] 1006[医学-中西医结合] 100706[医学-药理学] 100602[医学-中西医结合临床] 10[医学] 

基　　金：Project supported by the Committee of Science and Techno-logy of Guangdong Province No 001365 

主　　题：Agkistrodon acutus venom fibrinolytic enzyme hemorrhagic protease endothelial cells 

摘      要：AIM: To study the local hemorrhagic activity of a fibrinolytic enzyme (FIIa) from Agkistrodon acutus venom and its mechanism. METHODS: The local hemorrhagic activity was determined by subcutaneous injection on the back of mouse. The effects of FIIa on factor X, prothrombin, gelatin, and collagen were visualized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Platelet aggregation assays were performed in rat platelet- rich plasma (PRP). Human umbilical vein endothelial cells (HUVEC) were cultured and passaged in complete M199 medium. Cell viability and nuclear morphology change were determined by fluorescein diacetate (FDA) staining and Hoechst 33258 staining, respectively. RESULTS: The minimum hemorrhagic dose (MHD) of FIIa was 89 μg. In vitro, FIIa (0.25 g/L) degraded factor X, prothrombin, collagen, and gelatin, and dose-dependently (0.25, 0.50, 0.75, and 1.00 g/L) inhibited the platelet aggregation induced by ADP in rat PRP. When HUVEC in culture treated with FIIa, HUVEC showed detachment in a dose-dependent manner, but no apoptosis sign was observed. CONCLUSION: FIIahad local hemorrhagic activity, and the mechanism was related to the degradation of factor X, prothrombin, gelatin, and collagen, the inhibition of ADP-induced platelet aggregation, and inducement of HUVEC detachment.2Now in Beijing Universityof Chinese Medicine,Beijing 100029,China.3Correspondence to Dr Xiu-xia LIANG.