Exenatide Reduces Tumor Necrosis Factor-α-induced Apoptosis in Cardiomyocytes by Alleviating Mitochondrial Dysfunction.
Exenatide Reduces Tumor Necrosis Factor-α-induced Apoptosis in Cardiomyocytes by Alleviating Mitochondrial Dysfunction.
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艾塞那肽通过减轻线粒体功能障碍来减少肿瘤坏死因子-α 诱导的心肌细胞凋亡
DOI：
10.4103/0366-6999.170259
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发表时间：
2015-12-05
影响因子：
6.1
通讯作者：
Ge JB
中科院分区：
文献类型：
--
作者：
Cao YY;Chen ZW;Gao YH;Wang XX;Ma JY;Chang SF;Qian JY;Ge JB
肿瘤坏死因子-α(肿瘤坏死因子-α)在多种心脏疾病的进行性收缩功能障碍中起重要作用。肿瘤坏死因子-α的细胞毒作用部分是通过依赖于ROS和线粒体的细胞凋亡来实现的。胰升糖素样肽-1(GLP-1)或其类似物对心血管系统具有保护作用。本研究的目的是评价胰岛素样生长因子-1类似物埃塞那肽对肿瘤坏死因子-α诱导的体外培养心肌细胞氧化应激和细胞凋亡的影响。将分离的新生大鼠心肌细胞分为3组：对照组，细胞在正常条件下无干预培养；肿瘤坏死因子-α组，细胞与肿瘤坏死因子-α(40 ng/ml)共同孵育6、12、24 h而不加埃塞那肽；埃塞那肽组，在肿瘤坏死因子-α(40 ng/ml)刺激前30min用埃塞那肽(100nmol/L)预处理。用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法和流式细胞术检测细胞的凋亡，用特异的荧光探针检测ROS的产生和线粒体膜电位，用Western blotting检测各组的蛋白质水平。用流式细胞仪和原位末端标记法检测12 h和24 h时，艾塞那肽可显著减少心肌细胞的凋亡率，并抑制过量ROS的产生，维持心肌细胞的线粒体膜。此外，在埃克塞那肽处理的细胞中，细胞色素-c的释放减少，caspase-3的表达被切割，bcl2的表达增加，bax的活性降低。提示埃塞那肽对心肌细胞具有保护作用，可阻止肿瘤坏死因子-α诱导的心肌细胞凋亡，其抗凋亡作用可能与保护线粒体功能有关。
Tumor necrosis factor-α (TNF-α) plays an important role in progressive contractile dysfunction in several cardiac diseases. The cytotoxic effects of TNF-α are suggested to be partly mediated by reactive oxygen species (ROS)- and mitochondria-dependent apoptosis. Glucagon-like peptide-1 (GLP-1) or its analogue exhibits protective effects on the cardiovascular system. The objective of the study was to assess the effects of exenatide, a GLP-1 analogue, on oxidative stress, and apoptosis in TNF-α-treated cardiomyocytes in vitro. Isolated neonatal rat cardiomyocytes were divided into three groups: Control group, with cells cultured in normal conditions without intervention; TNF-α group, with cells incubated with TNF-α (40 ng/ml) for 6, 12, or 24 h without pretreatment with exenatide; and exenatide group, with cells pretreated with exenatide (100 nmol/L) 30 mins before TNF-α (40 ng/ml) stimulation. We evaluated apoptosis by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay and flow cytometry, measured ROS production and mitochondrial membrane potential (MMP) by specific the fluorescent probes, and assessed the levels of proteins by Western blotting for all the groups. Exenatide pretreatment significantly reduced cardiomyocyte apoptosis as measured by flow cytometry and TUNEL assay at 12 h and 24 h. Also, exenatide inhibited excessive ROS production and maintained MMP. Furthermore, declined cytochrome-c release and cleaved caspase-3 expression and increased bcl-2 expression with concomitantly decreased Bax activation were observed in exenatide-pretreated cultures. These results suggested that exenatide exerts a protective effect on cardiomyocytes, preventing TNF-α-induced apoptosis; the anti-apoptotic effects may be associated with protection of mitochondrial function.