活性氧在钙蛋白酶诱导血小板GPIbα酶切中的调控作用  

The Research Progress of Reactive Oxygen Species in the Regulation of Platelet Glycoprotein Ibα Ectodomain Shedding

作　　者：张平平[1] 张凤[1] 

机构地区：[1]安徽蚌埠医学院第一附属人民医院血液科,蚌埠233000

出　　处：《血栓与止血学》2016年第4期361-366,共6页Chinese Journal of Thrombosis and Hemostasis

摘　　要：目的探讨活性氧在血小板酶切中的调控作用。方法健康人洗涤血小板N-乙酰半胱氨酸(NAC)、二硫苏糖醇(DTT)、钙调蛋白酶抑制剂I(CII)和钙蛋白酶抑制剂II(MDL28170)和钙离子载体(A23187)凝血酶、钙蛋白酶活化剂(dibucaine)单独或联合处理。流式细胞仪分别进行血小板内ROS水平的变化和细胞膜表面GPIbα表达量的检测。处理好的血小板进SDS-PAGE电泳,NC膜转膜,用针对GPIbα的抗体SZ-2孵育,用胶片显影结果。SPSS软件对实验数据进行统计分析,不同组之间的比较采用one-way ANOVA进行统计分析,P<0.05表示有统计学差异。结果 1Dibucaine介导GPIbα酶切的同时,引起了血小板ROS水平的升高。2 DTT和NAC能够明显抑制Dibucaine诱导血小板内ROS水平,同时对Dibucaine诱导GPIbα胞外域的酶切产生抑制作用。3钙蛋白酶抑制剂显著降低A23187、凝血酶、Dibucaine诱导活性氧的产生。结论活化的血小板通过产生内源性活性氧诱导血小板GPIbα的酶切,该通路是依赖钙调蛋白酶的活化而实现的。Objective To study the the research progress of reactive oxygen species（ROS） in the reg- ulation of platelet glycoprotein Ibα ectodomain shedding. Methods N- acetyl- L- cysteine （ NAC ）, DTT, calpain inhibitors I（CII）, calpain inhibitors II（ MDL2817 ）or vehicle control （DMSO）were pre-incubated into washed platelets suspension separately, and then incubated with ）t23187, collagen, thromfiin and dibu- caine. The production of ROS and surface expression of GPIbct is detected by flow cytometers. The surface ex- pression of GPIbet ectodomain shedding is detected Western blot. Results ~ 1. dibucaine obviously induced ROS generation in platelets, and simultaneously, dibucaine incurred GPIbα shedding. 2. NAC and DTY both obviously reduced dibucaine-induced ROS production. Furthermore, NAC and DTT significantly.reduced dibu- caine-induced GPIbα ectodomain shedding. 3. ealpain inhibitors CI I and MDL obviously reduced A23187-, thrombin-and dibucaine-induced induced ROS production. Conclusion These data indicate that platelet ago- nists incur calpain activation resulting in ROS production in platelets.

关 键 词：血小板 GPIbα酶切 活性氧 钙蛋白酶 

分 类 号：R331.143[医药卫生—人体生理学]