Fig. 5

Improving mitochondrial dysfunction suppresses alcohol-induced hepatocyte pyroptosis. Hepatocytes were pretreated with 10 µM Drp1 inhibitor mdivi1 for 12 h, followed by stimulation with or without 40% alcohol. After stimulation, cells were collected for subsequent experiments. A The gene expression of mtDNA (ND1 and ND2) in hepatocytes was determined by qPCR. B Flow cytometry analysis of ROS by DCFH-DA in hepatocytes. C The protein expression of Mfn2, Drp1 and Fis1 in hepatocytes were measured by western blot. D The expression of GSDMD-N, NLRP3, ASC, cleaved Caspase 1, and pro-IL-1β in hepatocytes was determined by western blot. E The levels of IL-1β and IL-18 in the culture supernatants of hepatocytes were measured by ELISA kits.  F The LDH levels in the supernatants of hepatocytes were measured. G Fluorescence microscopy analysis of propidium iodide (PI) internalization expressed as PI positive cells percentages (%). Data are presented as mean ± SD. *p < 0.05, **p < 0.01