Jianpi Qingre Qushi Formula attenuates acute gouty arthritis through modulation of the Ras/Raf/MEK/ERK signaling pathway.

Abstract

The global incidence of acute gouty arthritis (AGA) continues to increase, and its pathogenesis is closely associated with complex inflammatory signaling pathways. Jianpi Qingre Qushi Formula (JQR) has shown favorable therapeutic efficacy in clinical practice; however, its key bioactive components and underlying molecular mechanisms remain unclear. This study integrated network pharmacology, molecular docking, and in vivo experiments to systematically investigate the potential mechanisms of JQR in AGA treatment. We screened active components of JQR and their potential targets, and collected AGA-related disease targets. A compound-target network was then constructed, followed by protein-protein interaction (PPI) analysis. Subsequently, GO (Gene Ontology) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were conducted on the overlapping targets. Molecular docking was performed to evaluate the binding affinities between core components and key signaling proteins. Additionally, a monosodium urate (MSU)-induced AGA rat model was established, and inflammatory responses and changes in key targets and signaling pathways were assessed using inflammatory scoring, histological examination, ELISA, immunofluorescence, western blotting, and quantitative PCR. JQR markedly alleviated inflammatory responses in AGA rats. Enrichment analyses indicated that the therapeutic effects of JQR were primarily associated with the Ras-dependent MAPK signaling pathway. Desacetylmatricarin, candletoxin A, dinatin, baicalein, and stigmasterol were identified as core active components and exhibited strong binding affinities to key proteins in the Ras/Raf/MEK/ERK signaling pathway. In vivo experiments further demonstrated that JQR notably reduced TNF-α and IL-1β levels, attenuated speck-like aggregation of the NLRP3 inflammasome, and downregulated the mRNA expression and protein levels of NLRP3, ASC, caspase-1, GSDMD, and IL-1β. In addition, JQR downregulated the protein level and mRNA expression of Ras, Raf, MEK, p-MEK, ERK, and p-ERK. These findings suggested that JQR exerted therapeutic effects on AGA via the coordinated regulation of multiple components and targets. This mechanism may be associated with the suppression of NLRP3 inflammasome activation and inflammatory responses through the Ras/Raf/MEK/ERK signaling pathway.