Formononetin and Rhein from Bitong Mixture Alleviate Rheumatoid Arthritis-Related Inflammation: An Integrated WGCNA,...

Area of Science:

• Pharmacology
• Computational Biology
• Immunology

Background:

• Bitong Mixture (BTM) is a traditional remedy for rheumatoid arthritis (RA).
• The precise active compounds and molecular mechanisms of BTM remain largely unknown.
• Understanding BTM's action is crucial for optimizing RA treatment.

Purpose of the Study:

• To identify the key molecular targets and active compounds of Bitong Mixture (BTM).
• To elucidate the anti-inflammatory mechanisms of BTM in rheumatoid arthritis (RA).
• To validate computational findings through in vitro experiments.

Main Methods:

• Integrated computational approaches including Weighted Gene Co-expression Network Analysis (WGCNA), network pharmacology, machine learning, and ROC analysis.
• Molecular docking and molecular dynamics simulations for target prioritization.
• In vitro validation using TNF-α-stimulated MH7A cells.

Main Results:

• JUN and PPARG were consistently identified as key targets across multiple analytical methods.
• Formononetin and Rhein demonstrated favorable binding affinity and stable interactions with identified targets.
• These compounds significantly reduced the expression of IL6 and MMP9 and suppressed fibroblast activation in vitro.

Conclusions:

• Bitong Mixture (BTM) likely exerts anti-inflammatory effects in rheumatoid arthritis (RA) via a regulatory network involving JUN and PPARG.
• Integrated computational and experimental evidence supports BTM's therapeutic potential.
• These findings provide a strong foundation for future in vivo studies and drug development.