粒细胞集落刺激因子对乳鼠缺氧心肌细胞的保护作用及机制探讨

目的研究粒细胞集落刺激因子(G-CSF)对缺氧心肌细胞的影响,探讨G-CSF发挥心肌细胞保护作用的分子机制。方法将心肌细胞分为3组,对照组、缺氧组和G-CSF组。流式细胞仪检测各组心肌细胞的存活率、凋亡率和坏死率。Northern blot法检测3组心肌细胞中Bcl-2、Bax、caspase-3 mRNA的表达,western blot法检测3组心肌细胞中细胞色素C(Cyt C)、信号转导与转录激活因子3(STAT-3)、caspase-3蛋白的表达。结果 6μg/LG-CSF开始发挥对缺氧心肌细胞的保护作用,150 μg/L保护作用最强,30μg/L和750μg/L保护作用相似。对照组、缺氧组和G-CSF组心肌细胞的存活率、凋亡率和坏死率差异有统计学意义。与缺氧组比较,G-CSF组Bcl-2mRNA表达增加,Bax、caspase-3 mRNA、Cyt C和caspase-3蛋白表达减少(P<0.01)。结论缺氧心肌细胞中,G-CSF可能通过线粒体途径发挥抗凋亡的作用。

Protective effect of granulocyte-colony stimulating factor on hypoxic cardiomyocytes and its mechanism

Objective To investigate the effect of granulocyte-colony stimulating factor(G-CSF)on hypoxic cardiomyoeytes and explore the possible protective molecular mechanism. Methods Cardiomyocytes were divided into three groups: normal cardiomyocytes, hypoxic cardiomyocytes and hypoxic cardiomyocytes treated with G-CSF. The survival,apoptotic and necrotic rates of eardiomyocytes were assessed by flow cytometry. Bcl-2, Bax and caspase-3 mRNA levels were determined by Northern-blot analysis and cytochrome C(CytC),STAT-3 and caspase-3 protein levels were determined by Western-blot analysis in each group. Results G-CSF at the concentration of 6 μg/L began to exert the protective effects, 150 μg/L of G-CSF was most effective,30 μg/L and 750 μg/L had similar protective effect. The survival, apoptotic and necrotic rates of three groups of cardiomyocytes were different significantly. Compared with the hypoxia group,the expression of Bcl-2 mRNA was upregulated and Bax, caspase-3 mRNA,Cyt C and caspase-3 protein were downregulated significantly in G-CSF group (P<0.01). Conclusions Hypoxic cardiomyocyte may be protected by activating chondriosome signaling pathway in G-CSF group.