Exploration of the molecular mechanism of Wufu Yin in the treatment of knee osteoarthritis based on network pharmacology and experimental validation.

Wufu Yin (WFY) exhibits significant clinical effectiveness in knee osteoarthritis (KOA) treatment, yet its therapeutic mechanisms are still unclear. This study aimed to explore the active ingredients and potential mechanism of WFY in the treatment of KOA. The network pharmacology-based approach was adopted to investigate the underlying mechanism of WFY in treating KOA. Molecular docking analysis was performed using Auto Vina software. An in vitro model of KOA inflammation was established by inducing chondrocyte cultures with interleukin-1 beta (IL-1β). Cell viability was quantified through the cell counting kit-8 assay, inflammatory cytokine levels were measured via ELISA, and protein expressions were assessed by Western blot analysis. A total of 225 active ingredients and 265 targets of WFY were identified, of which 88 were identified as potential targets against KOA. Enrichment analysis showed that these targets were associated with oxidative stress, cell proliferation and apoptosis, and inflammatory response, and were involved in the regulation of Th17 cell differentiation, IL-17 signaling pathway, tumor necrosis factor signaling pathway, and other signaling pathways. Topology analysis showed that PTGS2, NOS2, ESR11, PPARG, and MAPK14 had higher degree values and were key targets of WFY in the treatment of KOA. Molecular docking analysis showed that these key targets and active ingredients had low binding energies, indicating that they had potential binding activity. Furthermore, IL-1β-induced elevation of inflammatory cytokines, PTGS2 protein expression, and phosphorylated p38/p38 ratios in chondrocytes were significantly attenuated upon WFY intervention. Our study systematically elucidated the pharmacological basis and molecular mechanism underlying WFY's therapeutic effects in KOA, substantiating its ability to suppress inflammation and regulate PTGS2 expression and p38 phosphorylation.