【摘要】：目的探討抗霉素A(AMA)誘導(dǎo)血小板凋亡及其分子機(jī)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(xì)胞術(shù)檢測線粒體跨膜電位(ΔΨm)去極化、磷脂酰絲氨酸(PS)暴露、細(xì)胞內(nèi)活性氧(ROS)、線粒體ROS、P-選擇素表達(dá)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(yàn)中,先將洗滌血小板與線粒體靶向的ROS拮抗劑Mito-TEMPO預(yù)孵育,然后再與AMA孵育,流式細(xì)胞術(shù)檢測相關(guān)凋亡指標(biāo)。結(jié)果AMA劑量依賴性誘導(dǎo)血小板ΔΨm去極化、PS暴露、細(xì)胞內(nèi)ROS和線粒體ROS升高以及Caspase-3活化;但是AMA不能誘導(dǎo)血小板活化。線粒體靶向的ROS拮抗劑抑制AMA誘導(dǎo)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(jié)論 AMA能夠誘導(dǎo)血小板發(fā)生凋亡,線粒體ROS可能在AMA誘導(dǎo)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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