Palmitate (PA) exposure induces stress conditions featuring ROS accumulation and upregulation of p62 expression,resulting in autophagic flux blockage and cell apoptosis.Sulfuretin (Sul) is a natural product isolated from Rhus veiciflua Stokes;the cytoprotective effect of Sul on human hepatic L02 cells and mouse primary hepatocytes under PA-induced stress conditions was investigated in this study.Sul induced mitophagy by activation of p-TBK1 and LC3 and produced a concomitant decline in p62 expression.Autophagosome formation and mitophagy were assessed by the sensitive dual fluorescence reporter mCherry-EGFP-LC3B,and mitochondrial fragmentation was analyzed using MitoTracker Deep Red FM.A preliminary structure-activity relationship (SAR) for Sul was also investigated,and the phenolic hydroxyl group was found to be pivotal for maintaining the cytoprotective bioactivity of Sul.Furthermore,experiments using flow cytometry and weste blots revealed that Sul reversed the cytotoxic effect stimulated by the autophagy inhibitors 3-methyladenine (3-MA) and chloroquine (CQ),and its cytoprotective effect was almost eliminated when the autophagy-related 5 (Atg5) gene was knocked down.These studies suggest that,in addition to its antioxidative effects,Sul stimulates mitophagy and restores impaired autophagic flux,thus protecting hepatic cells from apoptosis,and that Sul has potential future medical applications for hepatoprotection.