Exploring the Therapeutic Mechanisms of Fangfeng Decoction for Rheumatoid Arthritis via Integrated Pharmacological, Lactylation, and Genetic Approaches.

ETHNOPHARMACOLOGICAL RELEVANCE: Rheumatoid arthritis (RA) is a chronic autoimmune disease that causes synovial inflammation and joint injury. Currently, safe multitargeted therapies for RA are lacking. Fangfeng Decoction (FFD), a 1,800-year-old traditional Chinese medicine, has been shown to relieve the symptoms of RA in clinical practice; however, its experimentally verified modern pharmacological mechanisms of action remain unclear.

MATERIALS AND METHODS: This study combined multi-omics, genetic analysis, and in vitro/in vivo experiments to explore FFD anti-rheumatoid arthritis (RA) effects: multi-omics and genetics identified FFD's core anti-RA targets/pathways, while subsequent assays (on MH7A cells and CIA BALB/c mice) verified these effects-including detection of apoptosis-related proteins and key signaling pathways.

RESULTS: Network pharmacology identified 142 FFD compounds (e.g., beta-sitosterol) targeting RA molecules (TNF-α and IL-6) and pathways (PI3K-AKT and NF-κB), which were validated by high-affinity molecular docking. Lactylation proteomics revealed that FFD reduced RA-related lactylation modifications (e.g., FTH1-K69la) and the lactate-H3K9la-NFATc2 axis. Mendelian randomization linked the FFD target HK3 to susceptibility to RA. In vitro, FFD inhibited RA synoviocyte proliferation in a dose-dependent manner, induced apoptosis (via Bcl-2/Bax/caspase-3), arrested the G1 phase, and suppressed PI3K/GSK-3β signaling. In vivo, FFD alleviated joint swelling, synovial hyperplasia, and pro-inflammatory markers (TNF-α, RF) in CIA mice.

CONCLUSIONS: FFD exerts experimentally verified multitarget, multi-pathway anti-RA effects, supporting its clinical potential as a complementary/alternative therapy for treating RA, especially in refractory cases, based on its modern pharmacological effects.