TFN modulates the TLR4/MAPK/NF-κB pathway via gut microbiota, thereby alleviating HUA-induced intestinal epithelial cells damage and enhancing UA excretion. To elucidate the phytochemical basis and therapeutic mechanisms of Tongfengning (TFN), a traditional herbal formula used to treat hyperuricemia (HUA), by integrating multi-omics strategies and functional validation. Ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS) was employed to identify the compounds in TFN. A HUA mouse model was induced using hypoxanthine (300 mg/kg) and potassium oxonate (250 mg/kg), followed by oral treatment with TFN at doses of 9.56, 19.11, 38.22 g/kg. Therapeutic efficacy was assessed via serum uric acid (UA) levels, renal function, intestinal barrier markers, and inflammatory mediators. Concurrently, 16S rRNA sequencing was utilized to analyze gut microbiota dynamics, and in vitro co-culture experiments demonstrated that TFN-modulated microbiota protect intestinal epithelial cells (IECs). Mechanistically, network pharmacology and RNA sequencing identified critical signaling pathways, which were subsequently validated by Western blotting. Multivariate statistical approaches were systematically applied to establish relationships between microbiota and HUA biomarkers. TFN significantly reduced serum UA, adenosine deaminase (ADA), xanthine oxidase (XOD) levels ( P < 0.01, 0.001), mitigated renal injury, and inhibited pro-inflammatory cytokines ( P < 0.05, 0.01, 0.001). TFN reduced serum levels of diamine oxidase (DAO) and D -lactate ( D -LA) ( P < 0.001), enhanced intestinal mucosal integrity, upregulated UA transporters, and enhanced intestinal UA excretion ( P < 0.05, 0.01, 0.001). TFN also restored the Firmicutes / Bacteroidetes ( F / B ) and mitigated microbiota dysbiosis. In vitro , co-culturing TFN-modulated gut microbiota with IECs directly protected against HUA-induced damage. Integrated network pharmacology with in vivo / in vitro transcriptomics identified the Toll-like receptor 4/mitogen-activated protein kinase/nuclear factor-κB (TLR4/MAPK/NF- κ B) axis as the key mechanism through which TFN exerts its therapeutic effects on HUA. Additionally, TFN decreased the abundance of Rhodobacteraceae, Halieaceae, Desulfocapsaceae, Anaerolineae, Akkermansia , Ilumatobacter , and Clostridium , while increasing that of Ruminococcus , Rikenellaceae, Prevotellaceae, Alistipes , Odoribacter , Erysipelotrichaceae, Eubacterium , Muribaculaceae, Monoglobus , Harryflintia , and Muribaculum , which were highly correlated with HUA markers or TLR4/MAPK/NF- κ B pathway. This study delineates a mechanistic network connecting the anti-HUA effects of TFN to intestinal barrier restoration, UA excretion, gut microbiota modulation, and TLR4/MAPK/NF- κ B pathway inhibition, while identifying key microbial signatures as potential therapeutic targets for HUA.
Guo et al. (Sun,) studied this question.