Inhibition of ROS/caspase-3/GSDME-mediated pyroptosis alleviates high glucose-induced injury in AML-12 cells

Diabetic liver injury (DLI) is a chronic complication of the liver caused by diabetes, and its has become one of the main causes of nonalcoholic fatty liver disease (NAFLD). The gasdermin E (GSDME)-dependent Pyroptosis signaling pathway is involved in various physiological and pathological processes; however, its role and mechanism in DLI are still unknown. This study was performed to investigate the role of GSDME-mediated Pyroptosis in AML-12 cell injury induced by high glucose and to evaluate the therapeutic potential of Caspase-3 inhibition for DLI. The results showed that high glucose activated Apoptosis by regulating the apoptotic protein levels including Bax, Bcl-2, and enhanced cleavage of Caspase-3 and PARP. Notably, some of the hepatocytes treated with high glucose became swollen, accompanied by GSDME-N generation, indicating that Pyroptosis was further induced by active Caspase-3. Moreover, the effects of high glucose on AML-12 cells could be partly reversed by a reactive oxygen scavenger (NAC) and Caspase-3 specific inhibitor (Z-DEVD-FMK), which suggests high glucose induced GSDME-dependent Pyroptosis in AML-12 cells through increasing ROS levels and activating Caspase-3. In conclusion, our results show that high glucose can induce Pyroptosis in AML-12 cells, at least in part, through the ROS/Caspase-3/GSDME pathway，and inhibition of Caspase-3 can ameliorate high glucose-induced hepatocyte injury, providing an important basis for clarifying the pathogenesis and treatment of DLI.