Objective: This study integrated network pharmacology and molecular docking to identify the active constituents, core targets, and potential mechanisms of Dunhuang Mofeng ointment in treating rheumatoid arthritis (RA). The therapeutic efficacy was validated using a collagen-induced arthritis (CIA) rat model. Methods: Bioactive compounds and target molecules of Dunhuang Mofeng ointment were identified using R language-based network pharmacological analysis. These were integrated with RA-related differential genes from the GEO database to construct regulatory and protein-protein interaction networks. Gene ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed to elucidate intervention mechanisms. Molecular docking simulations validated the conformational stability of core targets. A seven-week CIA rat model was established to assess therapeutic effects, including general health, joint swelling, limb deformities, serum inflammatory markers, and mRNA expression of key targets in joint tissues, thereby validating the network pharmacology predictions. Results: The core active ingredients were quercetin, apigenin, luteolin, aloe-emodin,and emodin. Primary therapeutic targets included STAT1, CDK4, CCND1, MCL1, FOS, CDKN1A and MYC, with anti-inflammatory effects mediated through pathways such as JAK-STAT, PI3K-Akt, and Toll-like receptor pathways. After 7 weeks of treatment, CIA rats showed significant improvements, including reduced joint swelling and deformity, decreased pro-inflammatory cytokines, and increased anti-inflammatory cytokine IL-10. MRNA expression of STAT1, CDK4, CCND1, MCL1, FOS, and MYC in joint tissues was significantly downregulated (P<0.05), while CDKN1A expression was upregulated (P<0.001). The medium-dose and Western medicine groups exhibited pronounced therapeutic effects. Conclusion: Dunhuang Mofeng ointment may exert therapeutic effects in RA by modulating pro-inflammatory cytokines and regulating key genes. These findings support its potential as an effective intervention for RA.