耐辐射奇球菌pprⅠ基因对MMC诱导H9C2心肌细胞损伤的保护作用

Protective effect of pprI gene of deinococcus radiodurans on injury of H9C2 cardiomyocytes induced by MMC

目的 探讨耐辐射奇球菌(DR)pprⅠ基因对丝裂霉素C(MMC)诱导H9C2心肌细胞损伤的保护作用及机制。方法 细胞分为空白对照组(不干预)、未转染组(MMC处理)、空质粒转染组(pCMV-C-Flag+MMC)与pprⅠ基因转染组(pCMV-C-Flag-pprⅠ+MMC)。CCK-8、抗氧化酶相关试剂盒、TUNEL、JC-1荧光探针和qRT-PCR测定细胞存活率、氧化应激、细胞凋亡率、线粒体损伤、凋亡途径相关基因mRNA的表达。结果 与空白对照组比较,未转染组细胞存活率、线粒体膜电位、B细胞淋巴瘤-2(Bcl-2)mRNA表达下降;氧化应激反应、细胞凋亡率及Bcl-2相关X蛋白(Bax)、半胱氨酸蛋白酶-9(Caspase-9)、Caspase-3、多腺苷二磷酸核糖聚合酶(PARP)mRNA表达增加(P<0.05)。与未转染组比较,pprⅠ基因转染组细胞存活率、线粒体膜电位、Bcl-2 mRNA表达增加;氧化应激反应、细胞凋亡率及Bax、Caspase-9、Caspase-3、PARP mRNA表达下降(P<0.05)。结论 耐辐射奇球菌pprI基因对丝裂霉素C诱导的H9C2心肌细胞损伤具有保护作用,其机制可能与激活线粒体凋亡途径有关。

Aim To investigate the protective effect and mechanism of pprⅠ gene of deinococcus radiodurans(DR) on injury of H9C2 cardiomyocytes induced by mitomycin C(MMC). Methods H9C2 cardiomyocytes were randomly divided into blank control group(no intervention), non-transfection group(MMC), empty plasmid transfection group(pCMV-C-Flag+MMC) and pprⅠ gene transfection group(pCMV-C-Flag pprⅠ+MMC). CCK-8, antioxidant enzyme related kit, TUNEL, JC-1 fluorescent probe and qRT-PCR were used to determine survival rate, oxidative stress, apoptosis rate, mitochondrial damage, and the mRNA expression of apoptosis pathway related genes. Results Compared with blank control group, the cell survival rate, mitochondrial membrane potential and B cell lymphoma-2(Bcl-2) mRNA expression were decreased in the non-transfected group;Oxidative stress reaction, apoptosis number and mRNA expression of Bcl-2 associated X protein(Bax), Caspase-9, Caspase-3 and poly(ADP-ribose) polymerase(PARP) increased(P<0.05). Compared with the non-transfected group, the cell survival rate, mitochondrial membrane potential and Bcl-2 mRNA expression in the pprⅠ gene transfected group increased;Oxidative stress reaction, apoptosis and the expression of Bax, Caspase-9, Caspase-3, PARP mRNA decreased(P<0.05). Conclusion The protective mechanism of pprⅠ gene of deinococcus radiodurans in mitomycin C induced H9C2 myocardial cell injury may be related to the activation of mitochondrial apoptosis pathway.