Xanthoxylin Protects Against Alcoholic Liver Injury via the EGFR/AKT Pathway: A Combined In Silico and In Vitro Study.

Alcoholic liver injury (ALI) represents a significant global health burden with limited therapeutic options. Xanthoxylin, a natural flavonoid compound, has demonstrated potential hepatoprotective properties, yet its underlying molecular mechanisms against ALI remain poorly elucidated. This study employed an integrated strategy combining network pharmacology, molecular docking, molecular dynamics (MD) simulations, and in vitro experimental validation to systematically investigate the protective mechanisms of xanthoxylin against ALI. Network pharmacology screening identified 52 intersection targets between xanthoxylin and ALI, with the top 10 core targets comprising ALB, PPARG, BCL2, PTGS2, ESR1, HIF1A, EGFR, HSP90AA1, GSK3B, and PARP1. GO enrichment analysis highlighted mitochondrion and mitochondrial outer membrane among the top 10 cellular component (CC) terms. KEGG pathway analysis revealed PI3K-Akt signaling within the top 10 pathways. Molecular docking suggested potential binding of xanthoxylin to the key targets. Subsequent MD simulations further confirmed the formation of stable complexes between xanthoxylin and EGFR, PPARG, and PTGS2. In vitro, xanthoxylin significantly ameliorated ethanol-induced HepG2 cell injury, attenuated TC and TG elevations, suppressed mitochondrial ROS accumulation, and enhanced SOD activity. Mechanistically, xanthoxylin upregulated HSP90, p-EGFR, EGFR, p-AKT, AKT, and PPARG protein expression, and suppressed the expression levels of PTGS2. Erlotinib, an EGFR inhibitor, reversed the cytoprotective effects of xanthoxylin. Xanthoxylin protects against alcoholic liver injury through regulating the EGFR/AKT pathway, with concurrent modulation of PPARG and PTGS2. These findings provide compelling evidence for xanthoxylin as a promising therapeutic candidate for ALI and establish a foundation for subsequent preclinical development.