大黄素通过激活线粒体caspase-8通路诱导L02细胞凋亡

Emodin induces apoptosis of L02 via caspase-8-mediated activation of the mitochondrial death pathway

目的:探讨大黄素对人正常肝细胞L02的毒性作用以及潜在的毒性作用机制。方法:通过Am-Blue法检测大黄素对L02细胞的毒性;使用DCFH-DA和JC-1探针检测细胞内活性氧水平与线粒体膜电位;使用Annexin V/PI双染色试剂,使用流式细胞仪检测凋亡细胞;凋亡相关蛋白caspase-8、caspase-9、caspase-3分别通过Ac-IETD-AFC,Ac-LEHD-AFC,Ac-DEVE-AFC荧光底物检测。结果:大黄素(20、40μmol/L)对L02细胞具有明显的细胞毒性,并且具有剂量依赖性;大黄素(20、40μmol/L)能显著升高细胞内ROS水平,使细胞线粒体膜电位降低。结论:大黄素通过激活线体caspase-8通路诱导L02细胞凋亡。

Objective： To investigate the effect and mechanism of emodin on cytotoxicity in L02 cells. Methods： Cell proliferation was assessed by Am-blue assay; The level of intracellular ROS was detected by the DCFH-DA fluorescent dye. The mitochondrial membrance potential was measured by the mitochondrial-specific lipophilic cationic fluorescent dye JC-1. Apoptotic cells were quantified by the Annexin V/PI double staining kit and analyzed by flow cytometry; Activities of caspase-8,-9 and-3 were measured by substrates Ac-IETD-AFC,Ac-LEHD-AFC,Ac-DEVE-AFC,respectively. Results： Emodin（20,40μmol/L） inhibited cell viability in a dose-and time-dependent manner. Emodin（20,40μmol/L） also elevated the production of intracellular ROS and the depolarization of mitochondrial membrane potential. Conclusion： Results demonstrate that the capsase-8 pathway is involved in emodin inducing apoptosis in L02 cells.