Network Pharmacology-Serum Metabolomics Integration Identifies Key Targets, Biomarkers and Mechanism of Sendeng-4 in Rheumatoid Arthritis Therapy.

Purpose: Rheumatoid arthritis (RA) is characterized by chronic inflammatory synovitis and immunometabolic dysregulation, necessitating safer multi-target therapies. Sendeng-4 (SD-4) is a traditional Mongolian medicinal formula composed of four botanical ingredients ( Methods: An integrative approach combining serum pharmacochemistry, network pharmacology, molecular docking, in vivo pharmacodynamics, and non-targeted serum metabolomics was employed. Absorbable constituents of SD-4 were identified by HPLC-Q-Exactive-Orbitrap-MS. Key targets and pathways were explored using network analysis, and therapeutic efficacy and metabolomic biomarkers were evaluated in a collagen-induced arthritis mouse model. Results: Twenty absorbable constituents were detected, with SRC, PIK3CA, and PIK3R1 emerging as key targets involved in PI3K-AKT and HIF-1 signaling. SD-4 treatment significantly reduced arthritis scores (by up to 45% in high-dose mice), paw thickness, and serum pro-inflammatory cytokines (TNF-α, IL-6, IL-1β decreased by 30-55%, all P < 0.05). Serum metabolomics identified 46 disease-associated metabolites reversed by SD-4, particularly involving tryptophan metabolism and glycolysis/gluconeogenesis. Correlation analyses suggest these metabolic changes are associated with modulation of inflammatory pathways. Conclusion: SD-4 alleviates arthritis in mice, likely through modulation of the PI3K/SRC network and partial rebalancing of glycolysis-tryptophan metabolic crosstalk, restoring immunometabolic homeostasis. These findings support the potential clinical application of SD-4 for RA and provide a mechanistic framework for its multi-target actions.