Oxypaeoniflorin Attenuates Alcohol‐Associated Liver Disease, Potentially Involving TEAD4 ‐Mediated Inflammatory Signaling Cascades

ABSTRACT This study investigates the hepatoprotective effects and underlying molecular mechanisms of Oxypaeoniflorin (Opae) against alcohol‐associated liver disease (ALD). An in vivo ALD mouse model was established by chronic feeding of Lieber‐DeCarli EtOH‐containing liquid diets combined with TEAD4 silencing is utilized to explore the protective function of Opae. Opae treatment effectively ameliorated alcoholic liver damage, accompanied by significant downregulation of hepatic P‐YAP1, TEAD1, and TEAD4 levels, as well as reduced expression of the pyroptosis‐associated proteins NLRP3 and IL‐1β. RNA sequencing analysis further identified the NOD‐like receptor/mitophagy/Hippo signaling axis as the core regulatory network mediating Opae‐induced hepatoprotection. Mechanistically, dual‐luciferase reporter assays in AML‐12 hepatocytes validated the direct binding of TEAD4 to the NLRP3 promoter. Both siRNA‐mediated TEAD4 silencing in AML‐12 cells and shRNA‐mediated hepatic TEAD4 silencing in mice markedly suppressed NLRP3 expression and alleviated liver inflammatory responses. Notably, TEAD4 depletion abrogated EtOH‐induced NLRP3 activation, confirming the essential transcriptional regulatory role of TEAD4 on NLRP3. In addition to the intrinsic regulatory effect on hepatocytes, Opae inhibits pyroptosis in LPS‐stimulated mouse peritoneal macrophages (MPMs). Conditioned medium collected from Opae‐treated MPMs significantly attenuated EtOH‐induced inflammatory injury in primary mouse hepatocytes. Collectively, Opae exerts prominent hepatoprotective effects in ALD by suppressing TEAD4‐NLRP3‐mediated pyroptosis.