雷公藤甲素诱导肝细胞L-02凋亡的机制

摘要

目的:探讨雷公藤甲素对人正常肝细胞L-02细胞的凋亡诱导作用及细胞线粒体呼吸链复合体抑制剂对细胞凋亡的影响.方法:采用Annexin V-FITC/PI双染色法检测雷公藤甲素对L-02细胞的凋亡作用,JC-1染色观察细胞线粒体膜电位的变化,线粒体呼吸链复合体抑制剂对细胞活性氧释放及细胞凋亡的影响.结果:流式细胞仪检测结果显示,雷公藤甲素对细胞增殖的抑制作用与浓度、时间呈正相关；JC-1染色可见经雷公藤甲素作用的细胞线粒体膜电位降低；加入细胞线粒体呼吸链抑制剂后,与单用雷公藤甲素组比较,雷公藤甲素+ rotenone组细胞活性氧释放水平显著降低(P＜0.05),同时抑制剂合用组细胞凋亡百分率显著降低(P＜0.05).结论:雷公藤甲素体外可诱导L-02细胞凋亡,机制可能是与公藤甲素上调经线粒体呼吸链复合体Ⅰ、Ⅲ产生的活性氧有关.%Objective： To investigat the apoptosis effects of triptolide on L-02 cells and the possible mito-chondrial respiratory chain-induced apoptosis mechanism. Methods; The apoptosis effects of triptolide on L-02 cells were evaluated using flow cytometry. JC-1 fluorescent staining was used to observe the mitochondrial membrane potential change of triptolide-treated cells. The effects of mitochondrial respiratory chain complex inhibitors on the release of ROS and cell apoptosis were analyzed. Results; Flow cytometry assay showed that the inhibition of triptolide on cell proliferation was positively correlated with the incubation concentration and time. JC-1 staining revealed the typical decrease of mitochondrial membrane potential. Compared with triptolide group, the ROS release level of triptolide + rotenone group significantly decreased(P <0.05) , while cell apoptosis was also significantly inhibited (P < 0. 05) in the presence of mitochondrial respiratory chain complex inhibitors. Conclusion; Triptolide probably inhibits the proliferation and induces the apoptosis of L-02 cells by up-regulating of ROS release from mitochondrial respiratory chain complex I and III.