Transcriptomic analysis combined with network pharmacology and molecular docking identifies NLRP3/SOD2 as potential targets of puerarin linked to macrophage polarization in lumbar disc herniation

Lumbar disc herniation (LDH) is a prevalent degenerative spinal disease with low back pain and neurological deficits. Puerarin, a major active component of leguminous plants, has anti-inflammatory and antioxidant effects, but its molecular mechanisms in regulating LDH remain unclear. GSE124272 transcriptomic data (8 LDH patients, 8 healthy controls) were integrated. Co-expression modules and hub genes were identified via WGCNA. LDH-associated genes were screened by combining limma-based differential analysis with CTD and GeneCard databases. Puerarin targets were predicted via CTD and SwissTargetPrediction, with overlapping genes with LDH selected for AutoDock-based molecular docking. An inflammatory model of THP1-M0 macrophages was established by LPS/ATP induction. The expressions of SOD2, NLRP3, and CD206 were detected by qPCR and Western blot, while the secretion levels of inflammatory cytokines were measured by ELISA. 27 co-expression modules and 969 hub genes were yielded via WGCNA. 36 key LDH-associated genes were identified by combining differential analysis and disease databases. SOD2 and NLRP3 were identified as overlapping genes between puerarin targets and LDH-associated genes. Favorable binding of puerarin to both proteins was demonstrated by molecular docking (NLRP3: −9.8 kcal/mol; SOD2: −7.1 kcal/mol). mRNA levels of SOD2 and M2 marker CD206 were significantly upregulated, while NLRP3 mRNA/protein levels were downregulated, and secretion of IL-6, IL-1β, and TNF-α was suppressed by puerarin. Puerarin may exert regulatory effects on LDH by targeting SOD2 and NLRP3, inhibiting inflammatory responses and promoting M2 macrophage polarization. This study provided novel potential targets and a theoretical basis for the clinical treatment of LDH. • WGCNA and network pharmacology identify NLRP3 and SOD2 as core targets of puerarin relevant to LDH. • Molecular docking verifies stable binding of puerarin to NLRP3 (-9.8 kcal/mol) and SOD2 (-7.1 kcal/mol). • In LPS/ATP-treated THP1 cells, puerarin modulates NLRP3 and SOD2, suppresses inflammation, and promotes M2 polarization.