线粒体对血小板膜蛋白GPIbα酶切的调控作用  

Regulatory Role of Mitochondria in the Shedding of Platelet Membrane Protein GPIbα

作　　者：赵丽丽[1] 戴克胜[1] ZHAO Li-Li;DAI Ke-Sheng(Jiangsu Institute of Hematology,National Clinical Research Center for Hematologic Diseases,NHC Key Laboratory of Thrombosis and Hemostasis,The First Affiliated Hospital of Soochow University,Collaborative Innovation Center of Hematology,State Key Laboratory of Radiation Medicine and Protection,Suzhou 215006,Jiangsu Province,China)

机构地区：[1]苏州大学附属第一医院,江苏省血液研究所,国家血液系统疾病临床医学研究中心,国家卫生健康委员会血栓与止血重点实验室,苏州大学省部共建放射医学与辐射防护国家重点实验室,江苏苏州215006

出　　处：《中国实验血液学杂志》2020年第5期1704-1709,共6页Journal of Experimental Hematology

基　　金：国家自然科学基金(81770117);江苏省科教强卫工程-临床医学中心资助项目(YXZXA2016002)。

摘　　要：目的:探讨线粒体对血小板膜蛋白GPIbα酶切的调控作用和机制。方法:分离健康志愿者外周静脉血血小板,选择破坏或保护血小板线粒体的功能的羰基氰-3-氯苯腙(CCCP)和环胞素A(CsA)、抑制膜蛋白GPIbα酶切的金属蛋白酶抑制剂GM6001、降低细胞内活性氧(ROS)水平的抗氧化剂NAC分别与洗涤血小板共同孵育,应用流式细胞术检测受刺激后血小板线粒体功能的变化对膜蛋白GPIbα酶切的影响。结果:CCCP诱导线粒体内膜两侧膜电位去极化,直接破坏线粒体的呼吸功能;血小板GPIbα的酶切检测发现GPIbα发生酶切,实验结果具有统计学意义。这提示当血小板线粒体功能受损伤时,会伴随出现GPIbα酶切现象;用CsA保护线粒体功能后,血小板GPIbα酶切现象被明显的抑制,实验结果具有统计学意义;而GM6001可以抑制GPIbα的酶切现象,但不影响线粒体的功能,实验结果具有统计学意义。这提示,血小板GPIbα的酶切在线粒体受到了调控。酶切调控酶ADAM17位于线粒体下游的信号通路上,用NAC抑制细胞内出现的氧化损伤,检测血小板受体的酶切变化发现,NAC可以有效地抑制血小板受体的酶切现象,并呈浓度梯度依赖,实验结果具有统计学意义。结论:线粒体功能异常引起血小板膜蛋白GPIbα发生酶切,其机制是源于异常功能线粒体导致胞内ROS代谢失衡,进而诱导酶切的发生。Objective:To investigate the role of mitochonaria in the regulation of platelet membrane protein GPIbαshedding and its mechanisms.Methods:The washed platelets were obtained from peripheral blood in healthy volunteers and co-incubated with mitochondrial inhibitor carbonyl cyanide m-chlorophenyl hydrazone(CCCP),mitochondrial protector cyclosporin A(CsA)or matrix metalloproteinases inhibitor GM6001.After the platelets was stimulated,the effect of mitochondria to the shedding in platelet membrane protein GPIbαwas detected by flow cytometry.Results:Depolarization of mitochondrial membrane potential and the respiratory function of mitochondrial could be induced and destroyed by the uncoupling agent CCCP.At the same time,the shedding of GPIbαwas detected out,and the result showed a statistical significance,which showed that the shedding of GPIbαcould be activated by the damaged of mitochondrial in platelets.After the mitochondrial was protected by CsA,the shedding of GPIbαwas inhibited significantly.GM6001 could only inhibited the shedding of GPIbα,but showed no inhibitation to the function of mitochondrial,which showed that the shedding of GPIbαwas regulated at the mitochondrial,and the regulatory enzyme of receptor shedding(ADAM17)was located in the pathway of downstream of mitochondria.After the oxidative damage in cells was inhibited by NAC,and the changes of GPIbαshedding was detected,the result showed that the GPIbαshedding could be inhibited by NAC,which showed a dose-dependent manner.Conclusion:The GPIbαshedding could be caused by abnormality function of metabolic,and the metabolic imbalance of ROS is caused by the abnormallity function of mitochondria.

关 键 词：血小板 血小板膜蛋白Ibα 活性氧 

分 类 号：R457.1[医药卫生—治疗学]