Exploring the potential mechanism of action of Wutou-Guizhi decoction in the treatment of rheumatoid arthritis through network pharmacology analysis.

As a widely recognized traditional Chinese medicine (TCM) decoction prescription in China, numerous studies have shown that Wutou-Guizhi decoction (WTGZD) exhibits significant therapeutic efficacy for Rheumatoid arthritis (RA). Nevertheless, the underlying molecular mechanisms have yet to be fully elucidated. This study aims to establish a database of active ingredients for WTGZD and identify RA-related target genes. The WTGZD-RA-Potential target gene network and protein-protein interaction network were constructed, followed by gene ontology (GO) analysis and functional enrichment analysis utilizing the Kyoto Encyclopedia of Genes and Genomes (KEGG). Cell proliferation was confirmed through CCK8 assay. Target gene identification was performed via real-time PCR using quantitative methods, and western blot analysis was conducted. In the course of this investigation, 95 active components of drugs and 34 targets associated with rheumatoid arthritis were identified. Through the utilization of network pharmacology analysis, we were able to identify a total of 17 essential active components of WTGZD and pinpoint 12 significant targets linked to rheumatoid arthritis (RA). Our findings suggest a consistent interaction between the key components of WTGZD and the critical targets associated with RA. Subsequent qPCR analysis revealed that stigmasterol, a principal constituent of WTGZD, exhibited inhibitory effects on the expression of various RA-related factors, such as TNF-α, IL-1β, MAPK8, MMP1, MMP3, and MMP9. Moreover, WTGZD effectively mitigated the increased protein expression of MMP-1 and MAPK8 induced by LPS stimulation, both of which are integral components of the IL-17 signaling pathway. These results suggest that WTGZD may play a significant role in the therapeutic intervention of rheumatoid arthritis by suppressing inflammatory immune responses.