小柴胡汤对人肝癌细胞Huh7细胞凋亡的影响

Effects of Xiaochaihu Tang on Apoptosis of Human Huh7 Cells in Liver Cancer

目的观察小柴胡汤对体外培养人肝癌细胞Huh7细胞凋亡的影响及相关机制。方法不同浓度小柴胡汤(0,0.5,1.0,1.5 mg/ml)处理人肝癌Huh7细胞24 h、48 h和72 h后,利用倒置显微镜观察细胞形态变化,MTT法检测细胞活力,Hoechst 33258染色观察不同浓度小柴胡汤对凋亡的影响,RT-PCR和Western blot检测Bax和Bcl-2的表达。结果不同浓度小柴胡汤作用Huh7细胞24 h能不同程度抑制细胞生长,MTT结果表明小柴胡汤干预Huh7细胞24 h、48 h和72 h,能显著抑制细胞活力,呈时间和剂量依赖性;小柴胡汤作用Huh7细胞24 h,细胞呈明显的凋亡特征;RT-PCR和Western blot结果表明,小柴胡汤促进Bax表达,抑制Bcl-2表达,与对照组比较差异有统计学意义。结论小柴胡汤促进Bax表达,抑制Bcl-2表达可能是其促进Huh7细胞凋亡的机制之一。

Objective To observe the effect of xiaochaihu tang on apoptosis in vitro cultivated Huh7 cells and the correlative mechanisms. Methods Xiaochaihu tang of different concentrations( 0,0. 5,1. 0,1. 5 mg / ml) was used for the treatment of Huh7 cells. In 24 h,48 h and 72 h,the inverted microscope was used to observe the morphological changes of cell,MTT assay was used to determine cell activity,Hoechst33258 was used to evaluate the impact on apoptosis and RT- PCR and western blot were used to detect the expressions of Bax and Bcl- 2. Results After treatment with xiaochaihu tang for 24 h,Huh7 cells were inhibited in growth. MTT results showed that the cell activity was inhibited significantly after the intervention for24 h,48 h and 72 h,indicating the time- dose dependence. In 24 h intervention,Huh7 cell showed apparent apoptotic characters. RT- PCR and western blot results showed that xiaochaihu tang promoted Bax expression and inhibited Bcl- 2 expression,indicating the significant differences as compared with the control group.Conclusion Xiaochaihu tang promotes Bax expression,inhibits Bcl- 2 expression,which may be one of the mechanisms for the promotion of Huh 7 apoptosis.