The Traditional Herbal Formula Tongluo Shizhi Fang Ameliorates Gouty Arthritis by Regulating the Inflammatory Microenvironment Through the TLR4-ABCG2 Signaling Pathway

Background Gouty arthritis (GA) is a common inflammatory joint disease characterized by monosodium urate crystal deposition, and effective therapies that simultaneously suppress inflammation and promote uric acid excretion remain limited. Tongluo Shizhi Fang (TSF), a traditional Chinese herbal formula composed of multiple medicinal herbs, has been widely used for the management of inflammatory disorders. Purpose This study was designed to evaluate the therapeutic effects of TSF against GA and to clarify its potential mechanisms of action. Materials and Methods In vitro cell-based experiments were performed. The Cell Counting Kit-8 assay was used to determine the optimal concentration (2 mg/mL) and treatment duration (24 h) of TSF. Subsequently, the pharmacological effects of TSF on fibroblast-like synoviocytes (FLS), inflammatory mediator production, and macrophage surface-marker profiles were evaluated. Finally, western blotting was performed to assess TSF-induced changes in the expression of key proteins involved in the toll-like receptor 4 (TLR4) signaling pathway and uric acid transport. Results TSF enhanced FLS viability and reduced apoptosis. It also increased scratch-wound closure in the co-culture model, although this assay reflects the combined effects of cell migration, cell number, and metabolic activity. TSF decreased interleukin (IL)-6 expression, increased IL-10 expression, and altered the surface-marker profile of macrophages. Because CD80 and CD206 alone do not define a complete functional polarization state, these findings were not interpreted as evidence that M1 polarization mediates the anti-inflammatory effect. Western blot analysis demonstrated that TSF downregulated TLR4, NF-κB p65, NLRP3, caspase-1, and ASC, while regulating PDZK1 and ABCG2 expression. Conclusion The traditional Chinese herbal formula TSF may regulate the inflammatory microenvironment through the TLR4-ABCG2 signaling pathway, thereby exerting potential therapeutic effects against GA. These findings provide preliminary pharmacological evidence supporting further investigation of TSF in GA.