Abstract: Objective To explore the role and mechanism of endoplasmic reticulum stress inhibitor GSK2606414 in preventing the apoptosis of AML12 cells induced by alcohol combined with palmitic acid. Methods C57BL/6 female mice were randomly divided into the control group（n=5）and model group（n=5）. The mouse models with alcoholic steatohepatitis were established by Lieber-DeCarli liquid diet. Immunohistochemistry, Western blot and RT-PCR were adopted to detect the expression levels of endoplasmic reticulum stress-related proteins ATF4 and CHOP. After normal liver cells of AML12 mice were treated with alcohol and palmitic acid for 24 h, the changes in the expression levels of Activating transcription factor-4（ATF4）and CCAAT/Enhancer binding protein homologous protein（CHOP）were detected. AML12 cells were pretreated with endoplasmic reticulum stress inhibitor GSK2606414, and then treated with alcohol and palmitic acid. Western blot was employed to detect the expression levels of ATF4 and CHOP, and TUNEL assay was utilized to determine the cell apoptosis. Results Serum ALT and AST levels and pathological results confirmed that the alcoholic steatohepatitis mouse model was successfully established. The expression levels of ATF4 and CHOP proteins in the model group were significantly higher than those in the control group（both P<0.001）. The expression levels of ATF4 and CHOP protein were significantly up-regulated after AML12 hepatocytes were stimulated with alcohol and palmitic acid（both P<0.001）. Compared with the non-pretreatment group, the expression levels of ATF4 and CHOP proteins were significantly inhibited and cell apoptosis was remarkably reduced in the AML12 cells pretreated with GSK2606414 inhibitor（both P<0.05）. Conclusion GSK2606414 can mitigate the AML12 cell apoptosis induced by alcohol combined with palmitic acid, probably due to the inhibitory effect upon the endoplasmic reticulum stress.