白藜芦醇对Hepa 1-6肝癌细胞凋亡和ROS的影响

Effects of Resveratrol on Apoptosis and ROS Production in Hepa 1-6 Hepatocarcinoma Cells

目的:观察白藜芦醇对小鼠肝癌Hepa 1-6细胞凋亡及活性氧(Reactive oxygen species,ROS)作用。方法:MTT比色法检测白藜芦醇对Hepa 1-6细胞增殖的影响;Hoechst 33258染色检测细胞凋亡形态;流式细胞术检测细胞凋亡;Western Blot法检测活化型Caspase-3;2',7'-二氯氢化荧光素乙二脂(DCF-DA)染色法结合荧光酶标仪检测细胞内ROS的产生。结果:与对照组比较,20、40、80μmol/L白藜芦醇作用24、48、72 h均可显著抑制肝癌Hepa1-6细胞增殖,呈一定的时间与剂量依赖性。不同浓度白藜芦醇作用48 h可促使Hepa 1-6细胞凋亡,呈现细胞凋亡形态改变,同时伴有活化型Caspase-3及ROS的产生。结论:白藜芦醇可以抑制Hepa 1-6细胞增殖,诱导肝癌细胞凋亡,可能与Caspase-3活化及细胞内ROS水平升高相关。

Objective：To observe the effects of resveratrol on apoptosis and ROS production in murine hepatocarcinoma Hepa 1-6 cells in vitro.Methods：Hepa 1-6 cells were treated with different dose of resveratrol（20 μmol/L,40 μmol/L,80 μmol/L）.Cell proliferation was detected with MTT assay at 24 h,48 h and 72 h.Hoechst 33258 staining was used to visualize apoptotic morphology.Cell apoptosis was detected by flow cytometry analysis.Activated caspase-3 was detected by western blot.Intracellular ROS production was observed by 2′,7′-dichlorofluorescin diacetate（DCF-DA） staining.Results：Compared with the control,upon treatment with 20~80 μmol/L resveratrol for 24 h,48 h or 72 h,the proliferation of Hepa 1-6 cells was significantly inhibited in a time-dose dependent manner.20~80 μmol/L resveratrol also induced apoptosis and apoptotic morphology change in Hepa 1-6 cells accompanied with caspase-3 activation and ROS generation.Conclusion：Resveratrol could inhibit cell proliferation,induce apoptosis in murine hepatocarcinoma Hepa1-6 cells,and the mechanism may associated with caspase-3 activation and ROS production.