活性氧参与血小板凋亡调控的研究  

Study of reactive oxygen species on the regulation of platelet apoptosis

作　　者：王秀娟[1,2] 张平平[1] 赵丽丽[1] 涂彧[2] 戴克胜[1] 

机构地区：[1]苏州大学附属第一医院、江苏血液研究所、卫生部血栓与止血重点实验室,215006 [2]苏州大学放射医学与防护学院

出　　处：《中华血液学杂志》2014年第6期511-514,共4页Chinese Journal of Hematology

基　　金：科技部973前期研究专项（2012CB526600）;国家自然科学基金重点项目（81130008）

摘　　要：目的 探讨活性氧在血小板凋亡中的调控作用.方法 健康人洗涤血小板用N-乙酰半胱氨酸（NAC）、辛可卡因和凝血酶单独或联合处理.用活性氧检测试剂盒结合流式细胞术检测血小板活性氧的产生和线粒体膜电位（△Ψm）的改变,用Western blot检测半胱氨酸天冬氨酸蛋白酶3（caspase-3）及抗凋亡蛋白Bcl-xL的表达情况.结果 ①NAC＋辛可卡因组活性氧荧光值低于单用辛可卡因组（0.66±0.11对1.06±0.08,P=0.001）,NAC＋凝血酶组活性氧荧光值低于单用凝血酶组（0.45±0.05对0.71±0.11,P=0.001）.②NAC＋辛可卡因组线粒体膜电位正常的血小板百分数高于单用辛可卡因组[（86.30±9.37）％对（13.52±3.01）％,P＝0,000],NAC＋凝血酶组高于单用凝血酶组[（93.00±3.03）％对（76.58±5.28）％,P＝0.000].③单用辛可卡因组caspase-3活化后的裂解片段明显多于DMSO对照组,NAC＋辛可卡因组caspase-3活化后的裂解片段明显减少.④NAC＋辛可卡因组Bcl-xL的表达较单用辛可卡因组明显增多,NAC＋凝血酶组Bcl-xL的表达高于单用凝血酶组.结论 NAC对辛可卡因或凝血酶引起的血小板凋亡具有明显的抑制作用,这种抑制作用主要是通过对活性氧的调控而实现的.Objective To study the effect of reactive oxygen species （ROS） on the regulation of platelet apoptosis.Methods Washed healthy human platelets were pre-incubated with N-caetyl-L-cysteine （NAC）,and then stimulated with dibucaine or thrombin.The production of ROS and depolarization of mitochondrial membrane potential （△ Ψm） were detected by flow cytometry.The activation of caspase-3 and expression of Bcl-xL were analyzed by Western blot.Results ①The average ROS fluorescence value of NAC＋dibucaine group was lower than that of dibucaine group （0.66±0.11 vs 1.06±0.08,P〈0.01）,while that of NAC＋thrombin group was also lower than that of thrombin group （0.45± 0.05 vs 0.71±0.11,P=0.001）.②The percentage of platelets with normal △ Ψm in NAC＋Dibucaine group was higher than that of dibucaine group [（86.30±9.37）% vs （13.52±3.01）%,P=0.000],while that of NAC＋thrombin group was also higher than that of thrombin group [（93.00±3.03）% vs （76.58±5.28）%,P=0.000].③Fragmentation generated by caspase-3 activation in dibucaine group was much more than that in DMSO control group,while the fragmentation in NAC ＋dibucaine group was significantly decreased.④The expression of anti-apoptosis protein Bcl-xL of NAC＋dibucaine group was significantly higher than that of the dibucaine group,while that of NAC ＋ thrombin group was also higher than that of thrombin group.Conclusion Through the regulation of ROS,NAC could inhibit the platelet apoptosis induced by dibucaine or thrombin.

关 键 词：活性氧 N-乙酰半胱氨酸 血小板 膜电位 线粒体 细胞凋亡 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]