论文部分内容阅读
目的在体外缺氧无血清条件下模拟心肌缺氧微环境,研究阿托伐他汀(Atorvastatin,ATV)对小鼠心肌干细胞(cardiac stem cells,CSCs)的抗凋亡作用及可能发挥作用的分子机制。方法分离培养小鼠CSCs,在缺氧无血清条件下用ATV处理12 h,应用流式细胞术和Hoechst 33342荧光染色观察细胞凋亡,并利用Western blotting方法检测通路蛋白单磷酸腺苷活化蛋白激酶(AMP-activated protein kinase,AMPK)、哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)的磷酸化水平。结果缺氧无血清条件引起明显的细胞凋亡,ATV处理组细胞凋亡率显著降低(p<0.05),且呈一定的浓度依赖性。ATV处理组AMPK磷酸化水平明显升高(p<0.05),加入AMPK阻断剂CompoundC可以显著阻断这种效应。结论在缺氧无血清条件下,一定浓度的ATV对CSCs具有显著抗凋亡作用,且AMPK/mTOR通路可能参与了该过程。
Objective To simulate myocardial hypoxia and microenvironment under hypoxic and serum-free conditions in vitro and to study the anti-apoptotic effects of Atorvastatin (ATV) on mouse cardiac stem cells (CSCs) and the possible molecular mechanisms . METHODS: Mouse CSCs were isolated and cultured. The cells were treated with ATV for 12 h under hypoxia and serum-free conditions. Flow cytometry and Hoechst 33342 staining were used to observe the apoptosis of mouse CSCs. Western blotting was used to detect the expression of ATP- (AMP-activated protein kinase, mammalian target of rapamycin (mTOR phosphorylation level. Results In hypoxia-free and serum-free conditions, apoptosis was significantly induced. The apoptosis rate in ATV-treated group was significantly decreased (p <0.05) and in a dose-dependent manner. The phosphorylation level of AMPK in ATV treatment group was significantly increased (p <0.05), and AMPK inhibitor CompoundC significantly blocked this effect. Conclusion Under hypoxia and serum-free conditions, a certain concentration of ATV has a significant anti-apoptotic effect on CSCs, and AMPK / mTOR pathway may be involved in this process.