Resveratrol ameliorates CCl4-induced acute liver injury by restoring Parkin-mediated mitophagy.

BACKGROUND

Overexposure to hepatotoxins is a frequent cause of acute liver injury (ALI). This study aimed to investigate the protective effect of resveratrol (RSV) against carbon tetrachloride (CCl₄)-induced ALI and its underlying mechanisms.

METHODS

In vitro, four hepatic cell lines (HepG2, Huh7, Hepa1-6, and AML12) were pretreated with RSV (10, 20, or 40 µg/mL) before CCl₄ exposure. Cell viability, reactive oxygen species (ROS) levels, and mitophagy-related protein expression were assessed. In vivo, mice were orally administered RSV (10-40 mg/kg) for 7 days prior to CCl₄-induced ALI. Liver histopathological, liver function, oxidative stress markers, and apoptosis were measured.

RESULTS

RSV significantly attenuated CCl₄-induced cytotoxicity and ROS overproduction in vitro. It activated the expression of PTEN-induced putative kinase 1 (PINK1)/parkin RBR E3 ubiquitin protein ligase (Parkin)-dependent mitophagy, as indicated by upregulated Parkin, PINK1 and LC3-II (microtubule associated protein 1 light chain 3 beta), and reduced p62 expression. In vivo, RSV ameliorated CCl₄-induced ALI, reducing histopathological damage and serum alanine aminotransferase (ALT) / aspartate aminotransferase (AST) levels. RSV enhanced hepatic antioxidant capacity and decreased lipid peroxidation. Furthermore, RSV reduced hepatocyte apoptosis and consistently activated the PINK1/Parkin mitophagy pathway in liver tissues. Both in vitro and in vivo, RSV with the high dose exhibits the most potent effects.

CONCLUSIONS

These findings demonstrate that RSV, particularly at the high dose exerts hepatoprotective effects against CCl₄-induced ALI, partly by activating PINK1/Parkin-dependent mitophagy and alleviating oxidative stress. This highlights mitophagy's critical role in RSV-mediated protection and offers novel insights into its therapeutic mechanisms.