Polydatin Alleviates APAP‐Induced Hepatotoxicity by Targeting C/EBPβ to Suppress the NLRP3 Inflammasome Signaling Pathway

ABSTRACT Objective To investigate the protective effects and underlying mechanisms of polydatin (PD), a natural Sirt1 agonist derived from Polygonum cuspidatum, against acetaminophen (APAP)‐induced acute liver injury (ALI), with a focus on the regulation of the NLRP3 inflammasome signaling pathway and associated inflammatory pathways. Methods Polydatin's hepatoprotective effects against APAP‐induced ALI were evaluated using a murine model and the AML12 cell line. Liver damage was assessed through histopathological staining and biochemical assays. The protective mechanisms of polydatin were investigated through network pharmacology and RNA sequencing. Protein levels related to inflammation are measured via Western blotting. The effect of polydatin on reactive oxygen species (ROS) was analyzed using immunofluorescence and flow cytometry. The interaction of polydatin with target proteins was analyzed through molecular docking, molecular dynamics simulations, surface plasmon resonance (SPR), and cellular thermal shift assay (CETSA). Results PD significantly ameliorated APAP‐induced liver injury by restoring redox homeostasis and suppressing the release of inflammatory factors. Transcriptome sequencing revealed a time‐dependent upregulation of NLRP3, a core component of the NOD‐like receptor signaling pathway, during APAP challenge. PD inhibited the NLRP3 inflammasome signaling cascade and reduced downstream inflammatory mediators. Mechanistically, PD decreased both the expression and acetylation of C/EBPβ, thereby blocking C/EBPβ‐mediated activation of NLRP3 inflammasome signaling pathway and attenuating hepatocyte inflammation and liver damage. Conclusion This study identifies a Sirt1/C/EBPβ/NLRP3 signaling axis through which polydatin exerts its hepatoprotective effects. These findings provide a mechanistic rationale for developing PD as a preventive or therapeutic agent for APAP‐induced ALI.