Abstract

Background Alcoholic hepatic fibrosis (AHF) ultimately leads to liver cirrhosis and even hepatocellular carcinoma. The aim of this study was to investigate the specific mechanism by which high mobility group protein B1 (HMGB1) regulated the activation of hepatic stellate cells (HSCs) in AHF. Methods CCK-8, EdU staining and flow cytometry assays were utilized to evaluate the viability, proliferation and apoptosis of LX-2 cells stimulated by ethanol. The effect of HMGB1 on cell glycolysis was assessed by cellular energy metabolism assays. The levels of Fe2+/Fe3+, ROS, GSH and MDA were detected to evaluate the effect of HMGB1 on ferroptosis. In addition, clinical liver tissue samples and an AHF mouse model were employed to further investigate the effect of HMGB1 on AHF. Results Ethanol stimulation significantly upregulated HMGB1 and HSC activation markers, enhanced glycolysis, and inhibited ferroptosis in LX-2 cells. Knockdown of HMGB1 suppressed ethanol-induced effects, including HSC activation and glycolysis promotion. However, these effects of HMGB1 knockdown were negated by an oxidative phosphorylation inhibitor. Furthermore, a ferroptosis inducer impeded ethanol-induced HSC activation. Overexpression of HMGB1 decreased the ferroptosis level in ethanol-stimulated LX-2 cells, which was reversed by a glycolysis inhibitor. These in vitro findings demonstrated that upregulated HMGB1 inhibited ferroptosis by enhancing glycolysis, thereby promoting HSC activation. In vivo validation data further confirmed that HMGB1 knockdown inhibited glycolysis, increased ferroptosis level, reduced HSC activation, and alleviated liver fibrosis in AHF mice. Ferroptosis inhibitor counteracted the impacts of HMGB1 knockdown on ferroptosis, HSC activation and liver fibrosis in AHF mice. Conclusion In summary, HMGB1 promoted the metabolism of HSCs towards glycolysis to inhibit ferroptosis, which eventually led to the activation of HSCs and the progression of AHF.