Ursolic acid mitigates hepatic ischemia-reperfusion injury by regulating the ALOX12/12(S)-HETE and PTGES/prostaglandin E2 axis via arachidonic acid metabolism pathway

Introduction: Hepatic ischemia-reperfusion injury (HIRI) is a critical contributor to poor prognosis after hepatobiliary surgery, yet effective therapeutic strategies remain limited. Ursolic acid (UA), a natural pentacyclic triterpenoid, has shown potential hepatoprotective properties, but its specific mechanism in alleviating HIRI remains unclear. This study aimed to investigate the therapeutic effect of UA on HIRI and elucidate its underlying molecular mechanisms, addressing the clinical need for targeted interventions. Methods: A murine HIRI model was established, and UA was administered to assess its impact on liver function. Serum levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured to evaluate hepatoprotection. Integrated network pharmacology and non-targeted metabolomics were employed to identify key pathways and metabolites involved in UA-mediated protection. Molecular docking was used to predict interactions between UA and target proteins. Experimental validation included assessment of the ALOX12/12(S)-HETE and PTGES/prostaglandin E2 axes. A cellular thermal shift assay (CETSA) was performed to confirm direct binding between UA and the PTGES protein. Results: UA administration significantly reduced serum AST and ALT levels in HIRI mice, confirming its protective role against liver injury. Integrated multi-omics analysis revealed the arachidonic acid metabolic pathway as a central hub for UA's protective effects. Key metabolites (prostaglandin H2 and prostaglandin E2) were markedly downregulated by UA, and UA downregulated the ALOX12/12(S)-HETE and PTGES/prostaglandin E2 axes. CETSA confirmed that UA directly binds to the PTGES protein, supporting its role as a target of UA. Conclusion: UA attenuates HIRI primarily by modulating the arachidonic acid metabolic pathway and inhibiting the ALOX12 and PTGES signaling axes. These findings highlight UA as a promising therapeutic agent for HIRI, with potential for clinical translation to improve outcomes in hepatobiliary surgery.