Isobavachin Ameliorates Acetaminophen‐Induced Liver Injury Through Modulation of CYP2E1‐Mediated Bioactivation and NRF2/PI3K/AKT Cytoprotective Pathways

Acetaminophen (APAP)-induced liver injury (AILI) remains a critical clinical challenge with limited therapeutic options. This research elucidates the mechanisms through which the natural flavonoid isobavachin (ISO) confers protection against APAP-induced liver injury. In vivo experiments demonstrated that ISO treatment significantly attenuated histopathological parameters, oxidative stress and inflammation, as indicated by serum ALT/AST, MDA, SOD/GSH, TNF-α/IL-1β levels. Mechanistically, ISO inhibited CYP2E1 expression and promoted NRF2 activation. Furthermore, integrated network pharmacology, transcriptomics, and molecular docking simulations revealed the PI3K/AKT signaling pathway as a potential targets of ISO, demonstrating high-affinity confirmed between ISO and PI3K (-8.6 kcal/mol) or AKT (-6.4 kcal/mol) respectively. western blotting assay confirmed that ISO activated PI3K/AKT signaling and upregulated downstream anti-apoptotic- protein BCL2. These findings collectively suggest that ISO alleviates AILI through multi-mechanisms: inhibiting CYP2E1-mediated APAP bioactivation, modulating of the NRF2/PI3K/AKT signaling, ISO attenuates oxidative damage, suppresses inflammatory activation, and inhibits apoptosis. The results suggest that ISO represents a potential therapeutic agent for managing APAP-induced hepatotoxicity.