PPARα/PGC-1α信号通路对阿霉素损伤H9c2心肌细胞能量代谢的调节

The energy metabolism regulation of PPARα/PGC-1α signal pathway in H9c2 cardiomyocytes treated with doxorubcin

目的:探讨过氧化物酶体增生物激活受体α(PPARα)/过氧化物酶体增殖物激活受体γ辅助激活因子1α(PGC-1α)信号通路对阿霉素损伤后H9c2心肌细胞能量代谢的调节。方法:将传代后生长良好的H9c2心肌细胞随机分为4组:空白对照组(C组)、阿霉素损伤组(DI组)、PPARα预激动组(PPA组)和PPARα预抑制组(PPI组),药物预处理后予以阿霉素建立心肌细胞损伤模型。细胞继续培养24h后,荧光定量RT-PCR方法检测PPARα、PGC-1α和P300基因表达变化,运用Western blotting方法测定PPARα、PGC-1α及P300蛋白表达情况,高效液相色谱法(HPLC)测细胞线粒体内的腺苷酸含量。结果:与对照组相比,阿霉素损伤组、PPARα预抑制组中PPARα、PGC-1α及P300的表达均下降(P<0.01),高能磷酸盐的含量均显著降低(P<0.01)。与阿霉素损伤组、PPARα预抑制组相比,PPARα预激活能够提高PPARα、PGC-1α及P300的表达(P<0.01),增加高能磷酸盐的含量(P<0.01)。结论:阿霉素能够通过减少PPARα/PGC-1α轴的表达降低H9c2心肌细胞内磷酸腺苷含量,影响H9c2心肌细胞的能量代谢。而PPARα/PGC-1α轴的预激活能够增加阿霉素损伤H9c2心肌细胞磷酸腺苷的含量,改善H9c2心肌细胞能量代谢。

Objective：To investigate the energy metabolism regulation of Peroxisome proliferator activated receptorα（PPARα）and Peroxisome proliferator activate coactivator（PGC-1α）pathway in H9c2 cardiomyocytes treated with DOX（Doxorubcin）.Method：H9c2 rat cardiomyocytes were randomly divided into four groups：Control group（Group C）,DOX group（Group DI）,PPARαpre-agonist group（PPA group）,PPARαpre-inhibit group（PPI group）.PPARα,PGC-1α and P300 protein levels and mRNA expression were analyzed by Western Blotting and Real-time PCR.The size of adenine acid pool were measured by High Performance Liquid Chromatography（HPLC）.Result：Compared with the Control group,the expression of PPARα,PGC-1α and P300,as well as high energy phosphate content were decreased in DI group and PPI group（P〈0.01）..Compared with DI group and PPI group,pre-activation of PPARαincreased the PPARα,PGC-1αand P300 mRNA and protein expression（P〈0.01）,and the content of high energy phosphate also increased（P〈0.01）.Conclusion：DOX can reduce the energy metabolism of H9c2 cardiomyocytes through impress the PPARα/PGC-1α signal pathway.Pre-activation of PPARα/PGC-1α signal pathway can increase the high energy phosphate in H9c2 cardiomycytes,and protect the energy metabolism from the DOX damage.