Unraveling the Mechanisms of Modified Si Miao San in Treating Gouty Arthritis: Network Pharmacology and Experimental Validation

ABSTRACT To explore the therapeutic mechanisms of Modified Si Miao San (mSMS) in the treatment of gouty arthritis (GA) using a combination of LC–MS/MS analysis, network pharmacology, and experimental validation. The pharmacokinetics of mSMS in rat serum were analyzed by LC–MS/MS, and its major active metabolites were identified. Network pharmacology integrated data from multiple databases (ChEMBL, TTD, YaTCM, and DisGeNET) to investigate the interactions between compounds, targets, and pathways. Experimental validation was performed using HE staining, PCR, Western blot, and serum biochemical assays. UHPLC‐QE‐MS identified 480 metabolites in the mSMS group, 414 in the drug‐containing serum group, and 411 in the control group. Venn analysis revealed 23 unique metabolites in the mSMS serum group. Network pharmacology analysis highlighted key biological processes, including extracellular response to stimuli and steroid metabolic processes. KEGG pathway analysis revealed potential therapeutic targets such as TLR4, NLRP3, and CASP‐1. Experimental results showed that mSMS alleviated inflammation in GA rats in a dose‐dependent manner. It also reduced SUA, BUN, and CREA levels, downregulated TLR4, P65, NLRP3, and CASP‐1 expression, and upregulated ARG‐1. mSMS exerts anti‐inflammatory and urate‐lowering effects in GA by modulating the TLR4/NF‐ κ B/NLRP3 signaling axis, enhancing ARG‐1 expression, and promoting uric acid metabolism.