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.
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,E-mail:
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2020 Oct;28(5):1704-1709. doi: 10.19746/j.cnki.issn.1009-2137.2020.05.046.
To investigate the role of mitochonaria in the regulation of platelet membrane protein GPIbα shedding and its mechanisms.
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 GPIbα was detected by flow cytometry.
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 GPIbα was detected out, and the result showed a statistical significance, which showed that the shedding of GPIbα could be activated by the damaged of mitochondrial in platelets. After the mitochondrial was protected by CsA, the shedding of GPIbα was 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 GPIbα was 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 GPIbα shedding was detected, the result showed that the GPIbα shedding could be inhibited by NAC, which showed a dose-dependent manner.
The GPIbα shedding could be caused by abnormality function of metabolic, and the metabolic imbalance of ROS is caused by the abnormallity function of mitochondria.
探讨线粒体在调节血小板膜蛋白糖蛋白Ibα（GPIbα）脱落中的作用及其机制。
从健康志愿者外周血中获取洗涤后的血小板，与线粒体抑制剂羰基氰化物间氯苯腙（CCCP）、线粒体保护剂环孢素A（CsA）或基质金属蛋白酶抑制剂GM6001共同孵育。刺激血小板后，通过流式细胞术检测线粒体对血小板膜蛋白GPIbα脱落的影响。
解偶联剂CCCP可诱导并破坏线粒体膜电位及线粒体的呼吸功能。同时，检测到GPIbα的脱落，结果具有统计学意义，表明血小板中线粒体受损可激活GPIbα的脱落。用CsA保护线粒体后，GPIbα的脱落明显受到抑制。GM6001仅能抑制GPIbα的脱落，但对线粒体功能无抑制作用，这表明GPIbα的脱落是在线粒体水平受到调节，且受体脱落的调节酶（ADAM17）位于线粒体下游通路。用N-乙酰半胱氨酸（NAC）抑制细胞氧化损伤后，检测GPIbα脱落的变化，结果表明NAC可抑制GPIbα的脱落，且呈剂量依赖性。
GPIbα的脱落可能由代谢功能异常引起，而线粒体功能异常导致活性氧（ROS）代谢失衡。
