Lithium Chloride Protects Aml-12 Cells Against Hypoxia/reoxygenation Induced Apoptosis In A Mitochondria-dependent Manner

Objective:Hepatic ischemia–reperfusion (I/R) injury occurs in a variety of clinical settings. This study investigates the effect of inhibitor of GSK-3β, LiCl on ischemia/reperfusion injury in AML-12 cells in vitro. The underlying mechanism is also investigated.Methods: To evaluate the effect of LiCl on ischemia/reperfusion induced cell death, Annexin V and MTT assay was detected. To investigate the mechanism of the hepatic protection by LiCl, the reactive oxygen species (ROS) level and the mitochondrial membrane potential (Δψm) were measured using an imaging technique; and western blotting was performed to examine cytochrome c release and JNK and GSK-3βactived in AML-12 cells.Results: We observed that LiCl protected AML-12 cells against H/R induced apoptosis and that LiCl inhibited ROS-dependent JNK activation and triggering the mitochondrial apoptotic pathway, caused a loss ofΔψm, the release of cytochrome c and caspase-3 and subsequent apoptosis. Furthermore, LiCl inhibited GSK-3βin H/R-treated AML-12 cells and inhibited the generation of ROS and downstream processes.Conclusions: Our finding reveal that LiCl can protect AML-12 cells against H/R induced apoptosis by preventing activation of ROS- JNK and mitochondria-mediated apoptosis pathway in a GSK-3β-inhibited manner. These data indicate that LiCl is a promising anti-apoptotic agent for improving AML-12 survival during hepatic ischemia–reperfusion (I/R) injury.