没食子酸对小鼠TM3细胞凋亡的影响

Effects of Gallic Acid on Apoptosis of Mice TM3 Leydig Cells

为了研究植物多酚类化合物没食子酸(Gallic acid,GA)对小鼠TM3睾丸间质细胞凋亡的影响,利用WST-1、JC-1、DAPI以及RT-PCR方法研究GA对体外培养的TM3细胞活力、线粒体膜电位、增殖凋亡相关基因表达的影响。WST-1结果显示,培养体系中添加0,20,100,200,400μmol/L的GA,处理24 h后可抑制TM3细胞活力,且呈剂量依赖性,与对照组相比较,20~400μmol/L处理组TM3细胞活力显著降低(P<0.05)。进一步研究发现,20μmol/L和400μmol/L GA显著抑制细胞增殖相关基因Cyclin B1及PCNA表达(P<0.05),促进细胞凋亡相关基因BAX表达(P<0.05),引起细胞线粒体膜电位降低,细胞核固缩,形成细胞凋亡小体。结果表明,体外培养体系中添加GA可以明显抑制TM3细胞活力,引起细胞细胞凋亡。

The present study was undertaken to elucidate direct effect of Gallic acid(GA),one of the phenolic compound,on mice TM3 Leydig cell apoptosis,using WST-1,JC-1,DAPI and real-time PCR techniques.The results showed that after adding different concentrations of GA(0,20,100,200 and 400μmol/L)to the culture system for 24 h,the viability of TM3 cells was down-regulated in a dose-dependent manner.Compared with control group,the viability of TM3 cells in 20 to 400μmol/L GA treatment groups was significantly decreased(P<0.05).GA inhibited the expression of cell proliferation related genes Cyclin B1 and PCNA mRNA(P<0.05),on the contrary,it promoted the expression of apoptosis related gene BAX mRNA,which resulted in mitochondrial membrane potential decrease,nuclear condensation and apoptotic body formation.In conclusion,GA had a significant inhibitory effect on the viability of TM3 cells,and thus promoted the apoptosis of TM3 cells.