【摘要】：目的探討抗霉素A(AMA)誘導血小板凋亡及其分子機制。方法取健康志愿者單采血小板,離心洗滌得到洗滌血小板,將洗滌血小板與不同濃度的AMA孵育后,流式細胞術檢測線粒體跨膜電位(ΔΨm)去極化、磷脂酰絲氨酸(PS)暴露、細胞內(nèi)活性氧(ROS)、線粒體ROS、P-選擇素表達和整合素αⅡbβ3活化;Western blot法檢測半胱氨酸天冬氨酸蛋白酶3(Caspase-3)的活化。在抑制試驗中,先將洗滌血小板與線粒體靶向的ROS拮抗劑Mito-TEMPO預孵育,然后再與AMA孵育,流式細胞術檢測相關凋亡指標。結果AMA劑量依賴性誘導血小板ΔΨm去極化、PS暴露、細胞內(nèi)ROS和線粒體ROS升高以及Caspase-3活化;但是AMA不能誘導血小板活化。線粒體靶向的ROS拮抗劑抑制AMA誘導的血小板ΔΨm去極化、PS暴露、Caspase-3活化以及線粒體ROS的生成。結論 AMA能夠誘導血小板發(fā)生凋亡,線粒體ROS可能在AMA誘導的血小板凋亡過程中起重要作用。
[Abstract]:Objective to investigate the apoptosis of platelets induced by anti-mycin A (AMA) and its molecular mechanism. Methods single platelet was collected from healthy volunteers and washed platelets were obtained by centrifugation. After incubating washed platelets with different concentrations of AMA, mitochondrial transmembrane potential (螖 蠄 m) depolarization and phosphatidyl serine (PS) exposure were detected by flow cytometry. The activation of cysteine aspartate protease 3 (Caspase-3) was detected by Western blot method with the activation of integrin 偽 鈪，

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