论文部分内容阅读
Aim: To investigate the mitochondria-initiated apoptosis pathway involved in Carbon tetrachloride (CCl4) hepatotoxicity in vitro. Methods: Several cytotoxicity endpoints, including WST-8 metabolism, lactate dehydrogenase leakage and morphological changes, were examined. The 5,5’-dithio-bis(2-nitrobenzoic acid)reaction was used to measure reduced glutathione level, and the malondialdehyde level was determined using the thiobarbituric acid assay. The release of cytochrome c and Bcl-XL was detected by West blot. Caspase-3 activity was measured using the fiuorogenic substrate Ac-DEVD-AMC. DNA fragmentation was used to evaluate cell apoptosis. Results: A time- and dose-dependent decrease in cellular glutathione content was observed, along with a concomitant increase in malondialdehyde levels following the application of CC14. Caspase 3 activity was stimulated at all doses of CCl4, with the most significant activation at 3 mmol/L.Cytochrome c was released obviously after CC14 treatment. A time-dependent decrease in Bcl-XL expression was observed. DNA fragmentation results revealed apoptosis and necrosis following CC14 treatment. Conclusion: Oxidative damage is one of the essential mechanisms of CC14 hepatotoxicity, which triggers apoptosis via the mitochondria-initiated pathway.