Network Pharmacology and Experimental Validation Unravel How Puerarin Improves Chronic Postoperative Pain.

Puerarin demonstrates therapeutic potential in alleviating chronic postoperative pain (CPSP) through multi-target mechanisms. While previous studies established its ferroptosis-inhibiting and anti-inflammatory properties via lipid peroxidation reduction and iron-mediated apoptosis regulation, its specific efficacy in CPSP remained unexplored. This study integrated network pharmacology with experimental validation using a skin/muscle incision-retraction (SMIR) rat model. Protein-protein interaction network analysis, Gene Ontology annotation, and KEGG pathway enrichment revealed puerarin's dual action pathway: modulating Th17 cell differentiation and regulating the HIF1 signaling axis. Molecular docking confirmed high-affinity binding between puerarin and five core targets: HIF1A, PTGS2, mTOR, RELA, and GSK3β. In vivo validation showed puerarin significantly elevated mechanical pain thresholds in SMIR rats while downregulating mRNA expression of these targets via qPCR. The compound's multimodal mechanism involves coordinated suppression of inflammatory signaling cascades and hypoxia-responsive pathways. These findings establish a robust methodology combining computational prediction with biological validation for herbal compound research.