Department of Clinical Laboratory Medicine.
Division of Urological Surgery.
Blood Coagul Fibrinolysis. 2021 Oct 1;32(7):434-442. doi: 10.1097/MBC.0000000000001054.
Adult chronic idiopathic thrombocytopenic purpura (cITP) is a chronic and usually life-long haemorrhagic disorder in which enhanced platelet destruction and weakened platelet production lead to thrombocytopenia. Platelets were isolated from blood samples collected from 40 adult patients with cITP and 40 healthy volunteers. Mitochondrial membrane potential (ΔΨm) and plasma membrane phosphatidylserine externalization were determined by flow cytometry, and activation of caspase-3 and expressions of Bax, Bak and Bcl-xL were analysed by western blotting. Flow cytometry showed increased mitochondrial depolarization and lower ΔΨm in platelets from adult patients with cITP. In addition, plasma membrane phosphatidylserine externalization was observed on platelets from adult patients with cITP, but rarely from healthy volunteers. Western blot analysis of platelet proteins revealed that, in adult cITP patients, caspase-3 was activated, which cleaved gelsolin and to release a 47-kDa fragment. Moreover, the expressions of Bax and Bak were elevated, and Bcl-xL was decreased markedly in platelets from adult patients with cITP. Our findings reveal, based on loss of mitochondrial membrane potential (Δψm), phosphatidylserine exposure, caspase-3 activation, enhanced expression of Bax and Bak, and attenuated expression of Bcl-xL, that platelet death in the pathogenesis of thrombocytopenia in chronic ITP in adults is apoptotic.
成人慢性特发性血小板减少性紫癜(cITP)是一种慢性且通常为终身性的出血性疾病,其特征为血小板破坏增强和血小板生成减弱导致血小板减少。从 40 名成人 cITP 患者和 40 名健康志愿者的血液样本中分离血小板。通过流式细胞术测定线粒体膜电位(ΔΨm)和质膜磷脂酰丝氨酸外翻,通过 Western blot 分析 caspase-3 激活以及 Bax、Bak 和 Bcl-xL 的表达。流式细胞术显示,成人 cITP 患者的血小板线粒体去极化增加,ΔΨm 降低。此外,在 cITP 成人患者的血小板上观察到质膜磷脂酰丝氨酸外翻,但在健康志愿者中很少见。血小板蛋白的 Western blot 分析显示,在 cITP 成人患者中,caspase-3 被激活,裂解凝胶蛋白并释放 47kDa 片段。此外,Bax 和 Bak 的表达上调,Bcl-xL 在 cITP 成人患者的血小板中明显减少。我们的研究结果表明,基于线粒体膜电位(Δψm)丧失、磷脂酰丝氨酸暴露、caspase-3 激活、Bax 和 Bak 表达增强以及 Bcl-xL 表达减弱,血小板死亡在成人慢性 ITP 血小板减少症的发病机制中是凋亡性的。
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