Integrated network pharmacology and experimental validation to elucidate the mechanism of Lysimachia paridiformis var. stenophylla Franch. in rheumatoid arthritis.

This study aimed to investigate the potential targets and mechanism of action of Chinese Miao Medicine Lysimachia paridiformis var. stenophylla Franch (ZhuiFengSan, LPSF) against rheumatoid arthritis (RA) using an integrated approach combining network pharmacology, molecular docking, and experimental validation. Components of LPSF were systematically collected from multiple databases, and RA-related targets were retrieved from disease databases. Cross-referencing identified 99 potential targets for LPSF against RA. A protein-protein interaction network revealed 5 core targets (TNF, IL6, AKT1, IL1B, and PTGS2), with TNF exhibiting the highest Degree value. Functional enrichment analysis indicated these targets were significantly associated with inflammatory responses and were notably enriched in the TNF signaling pathway. Molecular docking and molecular dynamics simulation confirmed strong binding affinities between TNF-α and several primary active components of LPSF, namely alternariol, galangin-3-methoxy, and alpinetin. Experimental validation demonstrated that LPSF significantly inhibited TNF-α-induced migration in MH7A cells and downregulated the mRNA and protein expression levels of key inflammatory factors (TNF, IL-6, IL-1β, and COX-2). Furthermore, LPSF inhibited TNF-α activity, increasing viability in L929 cells and reducing luciferase expression in an NlucP-NF-κB reporter gene system. In conclusion, this multi-dimensional study demonstrates that LPSF exerts its therapeutic effects against RA primarily by targeting TNF-α.