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目的探讨雷公藤甲素诱导人肝细胞L-02凋亡的机制。方法将L-02细胞分为对照组和雷公藤甲素50nM实验组,处理48h后,采用MTT法检测细胞存活率,流式细胞术检测细胞凋亡率、活性氧(ROS)的荧光强度,试剂盒检测细胞色素C的浓度以及半胱天冬氨酸蛋白酶9(Caspase-9)和Caspase-3的活性,Western blot检测细胞中B细胞淋巴瘤-白血病2(Bcl-2)和Bcl相关X蛋白(Bax)的表达。结果与对照组相比,实验组细胞存活率下降,细胞凋亡率上升,Bcl-2表达下调,Bax表达、细胞色素C浓度、ROS荧光强度、Caspase-9和Caspase-3活性均明显上调(P<0.05或P<0.01)。结论雷公藤甲素可以抑制L-02细胞的存活,通过激活线粒体凋亡途径促进L-02细胞凋亡,下调Bcl-2与Bax比率,促进细胞色素C释放,诱导ROS生成,进一步破坏线粒体膜,增加细胞色素C的释放,激活Caspase-9和Caspase-3介导的细胞凋亡通路,最终诱导细胞凋亡。
Objective To investigate the mechanism of Triptolide-induced apoptosis of human hepatocyte L-02. Methods L-02 cells were divided into control group and triptolide 50nM experimental group. After treated for 48h, cell viability was detected by MTT assay. Flow cytometry was used to detect the apoptosis rate, fluorescence intensity of reactive oxygen species (ROS) The concentration of cytochrome C and the activity of Caspase-9 and Caspase-3 were detected by Western Blot and Western blot. The expressions of Bcl-2 and Bcl-2 Protein (Bax) expression. Results Compared with the control group, the cell viability decreased, the apoptosis rate increased, the expression of Bcl-2 decreased, the expression of Bax, cytochrome C, ROS, Caspase-9 and Caspase-3 activity increased significantly P <0.05 or P <0.01). Conclusion Triptolide can inhibit the survival of L-02 cells, promote the apoptosis of L-02 cells by activating the mitochondrial apoptosis pathway, down-regulate the ratio of Bcl-2 to Bax, promote the release of cytochrome C, induce the production of ROS and further destroy the mitochondrial membrane , Increase cytochrome C release, activate caspase-9 and Caspase-3-mediated apoptosis pathway, and ultimately induce apoptosis.