Formononetin and Rhein from Bitong Mixture Alleviate Rheumatoid Arthritis-Related Inflammation: An Integrated WGCNA, Machine Learning and In Vitro Study.

Background

Bitong Mixture (BTM) is used clinically for rheumatoid arthritis (RA), but its active components and mechanisms remain unclear.

Methods

WGCNA, network pharmacology, machine learning, and ROC analysis were integrated to identify candidate targets. We used molecular docking and molecular dynamics simulations for prioritization. In vitro validation was performed in TNF-α-stimulated MH7A cells.

Results

Multiple analytical approaches consistently identified JUN and PPARG. Formononetin and Rhein showed favorable binding and stable interactions. Both compounds significantly reduced IL6 and MMP9 expression and suppressed fibroblast activation.

Conclusions

BTM may exert anti-inflammatory effects through a JUN- and PPARG-related regulatory framework, supported by integrated computational and experimental evidence, providing a foundation for further in vivo investigation.