Abstract

Drug-induced liver injury (DILI) represents a critical pharmacotherapeutic challenge requiring novel interventions. The hepatoprotective mechanisms of Gypenoside XLIX (GYP XLIX) against liver injury remain underexplored. This study establishes GYP XLIX as a hepatoprotective agent against diclofenac (DF)-induced liver injury through integrated experimental approaches. Male Wistar rats were used in this investigation and grouped randomly into four groups: the control group, the DF group (50 mg/kg), the GYP XLIX group (30 mg/kg), the DF + XLIX group, and the DF + silymarin group for 7 days. Human liver cell line L02 was used for further study. The analytical methods used included network pharmacology analysis, molecular docking, H&E staining, qPCR, enzyme activity assay, and Western blot. GYP XLIX significantly ameliorated DF-induced hepatic damage, as evidenced by reduced serum ALT, AST, and ALP levels, attenuated histopathological abnormalities, enhanced antioxidant capacity (elevated SOD and GSH, decreased MDA), and suppressed pro-inflammatory cytokine release. Mechanistically, GYP XLIX targeted the AKT/NLRP3 pathway, promoting AKT phosphorylation and inhibiting NLRP3 inflammasome activation, confirming its role in alleviating hepatotoxicity through antioxidative and anti-inflammatory mechanisms. In L02 cells, GYP XLIX exerts a protective effect against DF-induced liver injury through the AKT/NLRP3 axis, and this mechanism has been further confirmed. This work supports GYP XLIX as a promising multi-target therapeutic candidate for chemical hepatotoxicity, providing new insights for clinical prevention and treatment of DILI.