Ginsenoside Rg5 Ameliorates Ferroptosis in Ethanol-Induced Acute Liver Injury by Activating FTO-YTHDF2-Nrf2 Signal

Ginseng (Panax ginseng Meyer), one of the most widely known Chinese herbs, has been used in traditional medicine for centuries. Ginsenoside Rg5 (Rg5) can exert protective effects on the liver and activate the Nrf2 pathway. This study aims to investigate the mechanism of Rg5 alleviating acute liver injury (ALI), and the related mechanisms will be discussed. In vitro experiments, an ALI model was established using HepG2 cells. The DCFHDA, the JC-1, and the ferrous ion fluorescent probe detected the reactive oxygen species (ROS) level, the mitochondrial membrane potential change, and the iron ions level. The oxidative stress indexes were detected by biochemical analysis. Western blot was used to detect the m6A demethylation, Nrf2, and ferroptosis signaling pathways. For in vivo experiments, C57BL/6J mice were administered ethanol by gavage to establish the ALI model. In vitro, we observed that m⁶A methylation and FTO downregulation were involved in the ferroptosis process. Rg5 treatment alleviated ferroptosis after ethanol exposure, which was reflected by the decrease of intracellular iron content and ferroptosis-related proteins. The FTO knockdown exhibited ferroptosis-related proteins and YTHDF2 increase. The regulatory effect of FTO on ferroptosis was inhibited when Nrf2 was inhibited or knocked down. In vivo, HE staining revealed liver injury in the model group, with elevated liver-to-body weight ratios and serum ALT and AST levels. The Rg5 treatment has improved these phenomena. FTO, Nrf2, and anti-ferroptosis proteins were downregulated in the ethanol group, while the ferroptosis marker ACSL4 was upregulated. This study demonstrates that Rg5 alleviates ethanol-induced ALI by inhibiting ferroptosis via the FTO/YTHDF2/Nrf2 axis. Our work not only establishes a link between FTO-mediated m⁶A demethylation and ferroptosis but also provides new mechanistic insights into the hepatoprotective action of ginsenosides.