线粒体通透性转换孔道在脂多糖致H9c2心肌细胞凋亡中的作用

发布时间：2018-08-23 13:04

【摘要】：目的·观察脂多糖(LPS)刺激下心肌细胞线粒体通透性转换孔道(mPTP)开放状态、心肌细胞凋亡情况及环孢霉素A(CsA)和兰尼定(Rya)联合应用的抗凋亡作用。方法·将H9c2细胞分为对照组、LPS组、LPS+CsA组、LPS+Rya组、LPS+CsA+Rya组;分别检测mPTP的开放状态、细胞内及线粒体Ca~(2+)浓度、线粒体膜电位(ΔΦm),以及反映细胞凋亡的Hoechst染色,Bax、Bcl-2的mRNA表达,caspase 3活性,Bax及Bcl-2蛋白水平的变化。结果·LPS作用心肌细胞24 h后mPTP开放,可使细胞质Ca~(2+)荧光强度增高298%,线粒体Ca~(2+)荧光强度增高231%,约1/3细胞凋亡,caspase 3活性增加了约1倍,上调Bax mRNA表达(P=0.008);联合使用CsA与Rya可有效阻断由LPS引起的心肌细胞mPTP开放,降低LPS所致的细胞质和线粒体的Ca~(2+)浓度增高的幅度,维持正常的ΔΦm,减轻LPS所致的细胞凋亡,抑制caspase 3活性,可显著抑制LPS对心肌细胞Bax mRNA表达的上调作用。结论·mPTP在LPS所致心肌细胞凋亡中发挥重要作用,而CsA与Rya联合应用可有效缓解LPS所致的心肌细胞凋亡。
[Abstract]:Objective to observe the open state of mitochondrial permeability transition pore (mPTP) (mPTP) induced by lipopolysaccharide (LPS), the apoptosis of cardiomyocytes and the antiapoptotic effect of cyclosporin A (CsA) combined with lannitine (Rya). Methods H9c2 cells were divided into two groups: lipopolysaccharide (CsA) group, lipopolysaccharide (CsA Rya) group, open state of mPTP, intracellular Ca ~ (2) concentration and mitochondrial Ca ~ (2) concentration, respectively. Changes of mitochondrial membrane potential (螖桅 m),) and mRNA expression of Baxia Bcl-2 in mRNA and Bcl-2 protein. Results the opening of mPTP in cardiomyocytes induced by LPS for 24 h increased cytoplasmic Ca ~ (2) fluorescence intensity by 298, mitochondrial Ca ~ (2) fluorescence intensity increased by 231g, and apoptotic caspase 3 activity of about 1 / 3 cells increased by about one fold. The expression of Bax mRNA was up-regulated (P0. 008), the combination of CsA and Rya could effectively block the opening of mPTP induced by LPS, decrease the increase of Ca ~ (2) concentration in cytoplasm and mitochondria induced by LPS, maintain the normal 螖桅 m, and reduce the apoptosis induced by LPS. Inhibition of caspase 3 activity significantly inhibited the up-regulation of Bax mRNA expression by LPS. Conclusion mPTP plays an important role in cardiomyocyte apoptosis induced by LPS, and the combination of CsA and Rya can effectively alleviate the cardiomyocyte apoptosis induced by LPS.
【作者单位】：上海交通大学医学院附属第九人民医院检验科;上海交通大学医学院附属第九人民医院实验中心;
【基金】：上海市宝山区科学技术委员会项目(07-E-1)~~
【分类号】：R363

【参考文献】