The Synergistic Application of Triptolide and Celastrol Exhibits Efficacy and Enhancement in the Treatment of Rheumatoid Arthritis

Background Rheumatoid Arthritis (RA) is a persistent autoimmune disease. Triptolide (TPL) and celastrol (CEL) are both toxic components derived from traditional Chinese medicine Tripterygium wilfordii , which were proven to be potent candidates in the treatment of inflammatory and immunomodulatory. Objectives This study aims to explore the mechanism of action and efficacy of the combined application of TPL and CEL in the treatment of RA, as well as the advantages compared with their single use. Materials and Methods Network pharmacology predicted potential targets and pathways by analyzing interactions between RA and the 3D structures of TPL and CEL, and explored their combined effects. Cytotoxicity was assessed using Jurkat and RAW264.8 cell lines, and the dose ratio was validated. In vitro and in vivo studies further evaluated the synergistic therapeutic effects of TPL and CEL. Results Based on the topological importance of the “compound-target-pathway” network, inflammatory markers were identified as therapeutic targets of TPL and CEL, which play key roles in RA progression. TPL and CEL likely synergistically impact RA through these targets. Experimentally, their combination significantly inhibited inflammatory marker proliferation, mRNA/protein expression, and nuclear translocation in cells, outperforming individual treatments. The joint index confirmed their synergistic effect, aligning with network pharmacology predictions. In collagen-induced arthritis mice, TPL and CEL combined therapy markedly slowed RA progression, proving more effective together than alone. Conclusion The synergistic effect of TPL and CEL at a 1:160 ratio showed optimal efficacy for RA treatment with no significant toxicity. Both in vitro and in vivo studies confirmed its effectiveness over individual agents, attributed to nuclear factor kappa B and mitogen-activated protein kinase pathway inhibition.