抗霉素A诱导血小板凋亡的分子机制研究  

Study of molecular mechanisms on platelet apoptosis induced by antimycin A

作　　者：胡萍[1] 朱永明[1,2] 谢如锋[2] 王志成[3] 

机构地区：[1]华东师范大学生命科学学院,上海200062 [2]上海市血液中心血液工程研究室,上海200051 [3]复旦大学附属华山医院检验科,上海200040

出　　处：《安徽医科大学学报》2015年第11期1588-1593,共6页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81270650);上海市自然科学基金资助项目(编号:12ZR1429900)

摘　　要：目的探讨抗霉素A(AMA)诱导血小板凋亡及其分子机制。方法取健康志愿者单采血小板,离心洗涤得到洗涤血小板,将洗涤血小板与不同浓度的AMA孵育后,流式细胞术检测线粒体跨膜电位(ΔΨm)去极化、磷脂酰丝氨酸(PS)暴露、细胞内活性氧(ROS)、线粒体ROS、P-选择素表达和整合素αⅡbβ3活化;Western blot法检测半胱氨酸天冬氨酸蛋白酶3(Caspase-3)的活化。在抑制试验中,先将洗涤血小板与线粒体靶向的ROS拮抗剂Mito-TEMPO预孵育,然后再与AMA孵育,流式细胞术检测相关凋亡指标。结果AMA剂量依赖性诱导血小板ΔΨm去极化、PS暴露、细胞内ROS和线粒体ROS升高以及Caspase-3活化;但是AMA不能诱导血小板活化。线粒体靶向的ROS拮抗剂抑制AMA诱导的血小板ΔΨm去极化、PS暴露、Caspase-3活化以及线粒体ROS的生成。结论 AMA能够诱导血小板发生凋亡,线粒体ROS可能在AMA诱导的血小板凋亡过程中起重要作用。Objective To investigate the effects and molecular mechanism of platelet apoptosis induced by Antimycin A（ AMA）. Methods Washed healthy volunteers platelets were pre-incubated with different concentration AMA,and then depolarization of mitochondrial membrane potential（ ΔΨm）,phosphatidylserine（ PS） externalization assay,the surface expression of P-selectin,the activation of integrin αIIβ3 and the production of intracellular reactive oxygen species（ ROS）,mitochondrial ROS were detected by flow cytometry. The activation of Caspase-3was analyzed by Western blot. In inhibition experiment,washed platelets were pre-incubated mitochondrial ROS targeted antagonists Mito-TEMPO,and then stimulated with different concentration AMA. The apoptosis associated indicators were detected by flow cytometry. Results AMA dose-dependently induces depolarization of ΔΨm,PS exposure,Caspase-3 activation,and the production of intracellular ROS and mitochondrial ROS,but AMA does not induce platelet activation. Mito-TEMPO effectively reduces the depolarization of ΔΨm,PS exposure and the production of mitochondrial ROS which induced by the AMA. Conclusion AMA can induce platelet apoptosis,and mitochondria ROS may play an important role in the platelets apoptosis induced by the AMA.

关 键 词：血小板 凋亡 抗霉素A 线粒体 活性氧 

分 类 号：R331.143[医药卫生—人体生理学]