虎杖中白藜芦醇诱导肝癌细胞HepG-2凋亡及其对Bcl-2和Bax蛋白表达的影响

Apoptosis of HepG-2 Cell Induced by Resveratrol and Its Effects on Expressions of Proteins Bcl-2 and Bax

目的:探讨虎杖白藜芦醇对人肝癌HepG-2细胞增殖和凋亡。方法:采用微波辅助法提取虎杖中白藜芦醇,取对数生长期人肝癌HepG-2细胞,随机分为药物组和对照组,药物组用12.5,25,50,100μmol·L-1不同质量浓度的白藜芦醇作用人肝癌HepG-2细胞24,48,72 h后,采用MTT法检测虎杖白藜芦醇对人肝癌HepG-2细胞生长抑制作用;Hoechst 33342染色法荧光显微镜观察细胞的形态学变化,Western blot法检测不同浓度白藜芦醇作用HepG-2细胞48 h后B细胞淋巴瘤/白血病2(Bcl-2)蛋白和Bcl-2相关蛋白(Bax)的表达。结果:MTT实验结果表明,不同浓度白藜芦醇对人肝癌HepG-2的生长具有显著的抑制作用,呈现时间依赖性和剂量依赖性,当白藜芦醇质量浓度达到50,100μmol·L-1时,其对HepG-2细胞增殖抑制作用明显增强,与对照组相比,差异具有显著(P<0.01);荧光染色观察随着药物浓度增加,细胞凋亡数量增多,当50,100μmol·L-1白藜芦醇作用于人肝癌HepG-2细胞48 h可出现典型凋亡小体。Western blot法检测结果显示:随着白藜芦醇浓度的增加Bcl-2蛋白的表达逐渐降低,Bax蛋白的表达逐渐增加。结论:虎杖中白藜芦醇可以有效地抑制人肝癌HepG-2细胞增殖,诱导细胞凋亡,其诱导该细胞凋亡的作用机制可能是通过下调Bcl-2蛋白和上调Bax蛋白的表达。

Objective：To discuss the effects of resveratrol on the growth and cell apoptosis of human hepatoblastoma-derived HepG-2 cell line.Method：The resveratrol was extracted by microwave-assisted method and HepG-2 cells were exposed to resveratrol at the different concentrations.The hepatocellular carcinoma HepG-2 cells in logarithmic phase of growth were randomly divided into drug group and control group,with the concentration of 12.5,25,50,100 μmol · L-1 resveratrol in hepatocellular carcinoma HepG-2 cells.Intervention durations were 24,48,72 h accordingly.The viability of HepG-2 cells was measured by MTT.Morphotology of cell apoptosis was observed by Hoeschst 33342 fluorescence staining and fluorescent.The protein expression of Bcl-2 and Bax was analyzed by western blot.Result：Resveratrol inhibited the growth of cells and caused apoptosis significantly.The suppression was both in a time-and dose-dependent manner.The result of MTT showed that,at 50,100 μmol ·L-1 could inhibit the cell proliferation obviously compared with control group （P ＜ 0.01）.When HepG-2 cells were treated with the concentration of 50,100 μmol · L-1 resveratrol in 48 h,the typical apoptotic body can be observed under the fluorescence microscope.The expression of protein Bcl-2 was down-regulated and the expression of peotein Bax was up-regulated along with increase in resveratrol concentration.Conclusion：Resveratrol can inhibit the proliferation of HepG-2 cells in vitro and induce cell apoptosis.Resveratrol may induce apoptosis via down-regulating Bcl-2 and up-regulating of Bax in HepG-2 cells.