rheumatoid arthritis (ra) is a common systemic autoimmune disorder of unknown etiology, which threatens public health. The regulatory role of tripartite motif-containing 22 (TriM22) has been reported in multiple types of cancers and disease, but not in ra. The aim of the present study was therefore to elucidate the potential roles and underlying mechanisms of TRIM22 in fibroblast-like synoviocytes (FlSs) in ra. The Gene expression omnibus database was used to examine TRIM22 mRNA expression levels in synovial tissue samples of patients with RA and healthy controls. TriM22 and forkhead box c1 (FoXc1) mrna and protein expression levels in normal FLSs and RA-FLSs were assessed using reverse transcription-quantitative PCR (RT-qPCR) and western blotting, respectively. The Cell Counting Kit-8 assay was used to assess cell proliferation. cell apoptosis was analyzed using flow cytometry. The migratory and invasive abilities of ra-FlSs were assessed using Transwell assays.Western blotting was used to analyze the protein expression levels of apoptosis-related factors, MMP2, MMP9 and NF-κB signaling pathway-related proteins. inflammatory factors levels were assessed via ELISA and RT-qPCR. Furthermore, the JaSPar database, chromatin immunoprecipitation and the dual-luciferase reporter assays were used to determine the interaction between FoXc1 and the TriM22 promoter. The results of the present study demonstrated that TriM22 expression levels were significantly elevated in the synovial tissue samples of patients with RA and RA-FLSs. Moreover, FOXC1 was also significantly overexpressed in RA-FLSs. TRIM22 knockdown significantly reduced cell proliferation, migration, invasion and the inflammatory response, whereas cell apoptosis was significantly increased. Furthermore, the results demonstrated that FOXC1 may have positively mediated TRIM22 expression via binding to the TRIM22 promoter. Moreover, FOXC1 overexpression significantly reversed the outcome of TriM22 knockdown on the proliferation, apoptosis, migration, invasion and inflammation of RA-FLSs. FOXC1 overexpression also significantly reversed the inactivation of the nF-κB signaling pathway caused by TriM22 knockdown. in summary, the present study demonstrated that TriM22 was potentially activated via FOXC1, which contributed to the progression of RA via the NF-κB signaling pathway.

Background: Ermiao San, one Chinese medicine formula, has been widely used to treat rheumatoid arthritis(RA). Our previous study has demonstrated that Ermiao San is effective in treating RA. However, the pharmacological mechanisms remain unclear. Therefore, the purpose of this study was to decipher the potential mechanism of action of Ermiao San on rheumatoid arthritis (RA) by bioinformatics, network pharmacology, molecular docking, and molecular dynamics. Methods: Gene expression data (GSE77298) were from the GEO database. Differentially expressed genes (DEGs) were analyzed by R. The active ingredients of Huangbai (Phellodendron) and Cangshu (Atractylodes), two main constituents of Ermiao San, and their predicted target genes were retrieved from the Traditional Chinese Medicine Systems Pharmacology (TCMSP) platform. Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed using the overlapping genes between DEGs of the RA dataset and the predicted target genes of Ermiao San. The gene-gene interaction network was analyzed and visualized by Cytoscape. Molecular docking and dynamics simulations were performed to study the interaction between selected target genes (Chemokine ligand 2 (CCL2) and matrix metalloproteinase 1 (MMP1)) and active ingredients (quercetin and wogonin) of Ermiao San. Results: A total of 16 potential targets for Ermiao San were found, with their significantly enriched GO terms as cytokine-mediated signaling pathways, oxidoreductase activity, cell space, etc., and the most enriched KEGG terms as an IL-17 signaling pathway, rheumatoid arthritis, and NF-κB signaling pathway. CCL2 and MMP1 were identified and verified to be the targets of both quercetin and wogonin, the two active ingredients of Ermiao San, by molecular docking and molecular dynamics. Conclusion: Ermiao San may target CCL2 and MMP1 via its active ingredients to exert its therapeutic effects on RA.