Integrating Network Pharmacology, Molecular Docking, Molecular Dynamics Simulation, and Experimental Validation to Elucidate the Efficacy and Mechanism of Wanbi Hantong Yin in Rheumatoid Arthritis.

BACKGROUND

Wanbi Hantong Yin (WHY), a traditional Chinese medicine (TCM) formulation, has demonstrated therapeutic efficacy in rheumatoid arthritis (RA). However, its underlying molecular mechanisms remain largely unclear. This study combined bioinformatics analyses, including network pharmacology, molecular docking, and molecular dynamics (MD) simulation, with in vivo validation to elucidate the pharmacological mechanisms of WHY in the treatment of RA.

METHODS

A comprehensive WHY-Compound-Target-RA network was constructed to identify key bioactive compounds and their potential molecular targets. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed to determine the relevant biological processes and signaling pathways. Molecular docking and MD simulations were conducted to evaluate the binding stability and interaction strength between core compounds and key targets. The therapeutic efficacy of WHY was further validated in vivo using a collagen-induced arthritis (CIA) rat model.

RESULTS

Network pharmacology analysis identified quercetin, luteolin, kaempferol, and sitosterol as major active compounds, and STAT3, IL-6, TNF, JAK2, and IL-1β as principal targets. KEGG pathway analysis indicated that WHY exerted its anti-RA effects primarily through the regulation of Th17 cell differentiation and the TNF signaling pathway. Molecular docking and MD simulation confirmed stable binding interactions between the core compounds and key targets. In the CIA rat model, WHY significantly reduced the CD4+/CD8+ T cell ratio, increased Treg expression, and modulated Th17 differentiation. Furthermore, WHY decreased serum levels of TNF-α, IL-1β, IL-6, and IL-17, while increasing IL-10. It also suppressed oxidative stress, downregulated IL-6 mRNA expression, and inhibited activation of the JAK2/STAT3 signaling pathway.

DISCUSSION: RA is a chronic autoimmune disease characterized by progressive symmetric joint inflammation, leading to cartilage and bone damage, disability, and significant individual and societal burdens. Affordable, effective, and safe anti-RA treatments are urgently needed. WHY, a TCM prescription, has demonstrated efficacy in treating RA. However, the mechanism of WHY in treating RA remains unclear. This study explored the pharmacological mechanism of WHY in treating RA through network pharmacological analysis, molecular docking, MD simulation, and experimental validation. Our findings demonstrated that WHY modulated the immune response and reduced inflammation in RA by restoring T-cell homeostasis through the inhibition of the JAK2/STAT3 signaling pathway. This study provides novel insights into the mechanisms of TCM in RA intervention, fostering progress in new drug development.

CONCLUSION

This integrative study demonstrates that WHY restores T-cell homeostasis and alleviates inflammation in RA through inhibition of the JAK2/STAT3 pathway. These findings provide mechanistic insights into the immunomodulatory and anti-inflammatory effects of WHY, supporting its potential as an effective therapeutic agent for RA.