Ya-Jie-Sha-Ba Decoction Regulates Arachidonic Acid Metabolism to Treat Alcohol-Associated Liver Disease by Activating PPARα–CYP4A14 Axis: Insights From Multi-Omics Analysis and Experimental Validations

Abstract

Ya-Jie-Sha-Ba decoction (YJSB), a traditional Dai medicinal formulation, has been widely used by the Dai ethnic group and has shown efficacy in alleviating alcohol-associated liver disease (ALD). However, the molecular mechanism of YJSB in the management of ALD remains unclear. Building on evidence that YJSB ameliorates ALD, we combined transcriptomics and untargeted metabolomics to elucidate the molecular mechanisms responsible for its anti-ALD effects. We established an ALD mouse model and treated animals with YJSB to evaluate its therapeutic effects. We then performed integrated transcriptomic and untargeted metabolomic analyses to identify pathways and molecules associated with YJSB treatment. Using UPLC-Q-TOF-MS, we identified YJSB prototype components present in plasma. We screened the major plasma-detected constituent for hepatoprotective activity in an in vitro model of alcohol-induced hepatocyte injury. Finally, we validated the molecular target of this active component using molecular docking, cellular thermal shift assay (CETSA), drug affinity responsive target stability (DARTS), and pharmacological inhibition assays. The results showed that YJSB markedly reduced hepatic injury and lipid accumulation in ALD mice and simultaneously decreased oxidative stress and inflammation. Integrated multi-omics analysis identified arachidonic acid metabolism as a potential pathway mediating YJSB's protective effects. In particular, YJSB robustly increased the expression of CYP4A14, a key enzyme in this pathway. We identified methyl palmitate as the major active component of YJSB that mitigated alcohol-induced hepatocyte injury in vitro. Methyl palmitate decreased oxidative stress and enhanced CYP4A14 expression in injured hepatocytes. Molecular docking demonstrated a preferable binding affinity between methyl palmitate and PPARα. CETSA and DARTS further validated this interaction by showing that methyl palmitate increased the thermal stability and proteolytic resistance of PPARα. Importantly, pharmacological inhibition of PPARα completely abolished the hepatoprotective effects of methyl palmitate in vitro. In conclusion, YJSB effectively reduces liver injury in ALD mice. Mechanistically, its active component methyl palmitate activates PPARα, thereby upregulating CYP4A14, reshaping arachidonic acid metabolism, and attenuating alcohol-induced oxidative stress in hepatocytes.