视黄酸对乳鼠心肌细胞凋亡及凋亡相关因子表达的影响

Influence of retinoic acid on apoptosis and related genes expression in suckling rat myocardial cell

目的探讨视黄酸对乳鼠心肌细胞凋亡及凋亡相关基因、蛋白表达的影响。方法原代培养Balb/c乳鼠心肌细胞,并纯化获得心肌细胞。用不同浓度视黄酸(0.5、1.0、2.0、4.0μmol/L)进行处理,24h后倒置显微镜下观察各组心肌细胞的形态、搏动情况;流式细胞仪检测心肌细胞凋亡率;RT-PCR检测凋亡相关基因———Fas,Fasl mRNA的表达。Western blot测定乳鼠心肌细胞Fas,Fasl蛋白的表达。结果不同浓度的视黄酸对心肌细胞的影响不同。与对照组相比,低浓度组(0.5、1.0μmol/L)对心肌细胞形态、搏动影响不明显,Fas、Fasl mRNA及蛋白表达增加,差异无统计学意义(P>0.05);当视黄酸的浓度大于2.0μmol/L时,心肌细胞损伤明显,搏动力减弱、凋亡增加,Fas、Fasl mRNA及蛋白表达均明显增高(P<0.05)。结论高浓度视黄酸对心肌细胞有明显的抑制及促凋亡作用,与Fas,Fasl介导的死亡受体信号转导途径密切相关。

Objective To explore the influence of retinoic acid（RA） on apoptosis and related genes and apoptosis protein expression in suckling rat myocardial cell. Methods Balb/c suckling rat myocardial cells were performed the original generations cultivation, purified for obtaining myocardial cells and treated by different concentrations of retinoic acid（0.5,1.0,2.0,4.0 μmol/L）. After 24 h,the morphology and pulse status of myocardial cells in various groups were observed under phase contrast microscope. The apoptosis rate of myocardial cells was detected with flow cytometry. The apoptosis-related gene Fas, Fasl mRNA expression were determined by RT-PCR. Western blot technique was adopted to detect expression of Fas, Fasl protein in suckling rat myocardial cells apoptosis. Results Different concentrations of RA had different influence on myocardial cells. Compared with the control group, the low concentration group（0.5,1.0 t^mol/L） had no obvious influence on the morphology and pulse of myocardial cells, the expression of Fas and Fasl mRNA / protein was increased without statistical difference（P^0.05）. However,in the RA concentrationS2.0 μmol/L, the damage of myocardial cells was obvious, pulse was weakened, apoptosis was increased and Fas, Fasl mRNA and protein expression were significantly increased（P〈0.05）. Conclusion High concentration of RA has obvious inhibiting effect on myocardial cells and promotes apoptosis, which is closely related with Fas, Fasl mediated death receptor signal transduction pathways.