Unraveling the Gut Microbiota-Mediated Mechanism of Guogong Jiu for Rheumatoid Arthritis Treatment through Network Pharmacology and Metabolomics

INTRODUCTION: Rheumatoid Arthritis (RA) is a chronic autoimmune inflammatory disease. Guogong Jiu (GGJ) is a classical traditional Chinese medicine formula that is widely used in clinical treatment of rheumatoid arthritis. Nevertheless, the gut microbiota and metabolic mechanisms have not been fully studied. The objective of this study was to elucidate the gut microbiota-mediated mechanisms by which GGJ alleviates RA using network pharmacology, metabolomics, and experimental approaches. METHODS: A rat model of collagen-induced arthritis was established to assess anti-arthritic effects systemically. The arthritis index, histopathology, inflammatory cytokines, and gut microbiota analysis (16S rRNA) were assessed for their effects. The serum and fecal metabolomics were done using UHPLC-Q-Exactive MS/MS. Moreover, network pharmacology and secondary metabolome analysis were used to identify the active herbal components, potential targets, and pathways, which were validated in Rheumatoid Arthritis Fibroblast-Like Synoviocytes (RA-FLS) as well. RESULTS: GGJ improved the symptoms of Rheumatoid arthritis. GGJ affected pathways involved in amino acid, lipid, and energy metabolism. Moreover, the assessment of gut micro-biota revealed that GGJ helped restore microbial equilibrium by augmenting beneficial bacteria populations, including Lactobacillus and Alloprevotella, alongside diminishing Prevotella abundance. An integrated analysis identified NF-κB, MAPK, and NRF2 as key targets, which were subsequently validated at the cellular level. DISCUSSION: The multi-omics integration reveals that GGJ exerts its therapeutic effects through coordinated regulation of the gut-joint axis, involving microbiota restoration, metabolic reprogramming, and signaling pathways. These findings provide a mechanistic basis for the clinical application of GGJ in RA. CONCLUSION: GGJ is effective in RA through multi-target effects involving modulation of inflammatory signaling pathways, mediation of metabolic reprogramming, and restoration of gut microbiota. The clinical application of GGJ for RA is therefore scientifically supported.