黄芩苷诱导人肝癌HepG-2细胞凋亡作用观察

目的研究黄芩苷对人肝癌HepG-2细胞凋亡的影响,并探讨其作用机制。方法以不同浓度的黄芩苷与人肝癌HepG-2细胞共同培养,采用MTT法测定细胞增殖抑制率;采用Hoechst33258/PI染色法在荧光显微镜下观察凋亡细胞形态学变化;采用TUNEL法检测细胞凋亡率;采用Western blot法检测细胞凋亡相关蛋白Caspase-9、Caspase-3和Bcl-2表达的变化。结果在25~100μg/ml的药物浓度作用下,细胞增殖被抑制。药物浓度在50μg/ml下作用48h,细胞抑制率达50.63%,药物浓度在75μg/ml下作用48h细胞,抑制率达77.62%。抑制率呈浓度和时间依赖性;药物浓度在50μg/ml时作用48h,可见HepG-2细胞皱缩、细胞核碎裂成碎片,呈现典型的凋亡改变;随着黄芩苷作用浓度的增加,细胞凋亡率增高(P<0.05);随药物浓度的增加,Caspase-9和Caspase-3蛋白表达量呈增加趋势,而Bcl-2表达减少。结论黄芩苷能通过诱导细胞凋亡进而抑制人肝癌细胞增殖,其诱导凋亡机制可能与线粒体通路有关。

Inhibition of apoptosis of HepG-2 cells by bacalin in vitro

Objective To investigate the inhibition of apoptosis of HepG-2 cells by bacalin in vitro.Methods Human hepatocellular carcinoma cells,HepG-2 were cultured with different concentrations of baicalin and the inhibitory rate of cell proliferation was calculated with MTT assay;Morphology changes were detected by Hoechst33258/PI double fluorescence coloration;The apoptosis rate was detected by TUNEL;the Caspase-9,Caspase-3 and Bcl-2 protein expression were detected by Western blot.Results The cell proliferation was inhibited by baicalin at concentration of 25 to 100μg/ml;the inhibitory rate in 48h was 50.63% at 50μg/ml and 77.62% at 75μg/ml,and the inhibitory effect was in the concentration and time-dependent manner;there was a typical apoptosis at 50μg/ml in 48h,showing cell shrinkage and fragmentation of cell nucleus;the caspase-9 and caspase-3 expression increased and Bcl-2 decreased as the concentration of baicalin increased.Conclusion Baicalin can inhibit proliferation of HepG2 cells by inducing apoptosis and the mechanisms might be related to the mitochondrial pathway.