Exploring the mechanism of action of Qufengzhitong pills in treating rheumatoid arthritis via the PI3K/AKT, NF-κB, and MAPK pathways

OBJECTIVES: To elucidate the therapeutic mechanism of Qufengzhitong Pills (QFZTP) in the treatment of rheumatoid arthritis (RA). METHODS: Blood-absorbed active components of QFZTP and its potential therapeutic targets for RA were screened via liquid chromatography-tandem mass spectrometry (LC-MS/MS) and network pharmacology. Core targets were analyzed using protein-protein interaction networks, with key pathways identified via Gene Ontology/Kyoto Encyclopedia of Genes and Genomes enrichment. Molecular docking was performed for major components and core targets. An IL-1β-induced MH7A cell-based RA model was established; CCK-8, ELISA (IL-6/TNF-α), wound-healing/Transwell, flow cytometry, EdU, and western blot (PI3K/AKT/NF-κB/MAPK pathways) were used to assess cell viability, cytokines, migration, apoptosis, proliferation, and protein expression. KEY FINDINGS: LC-MS/MS identified 92 active components in rat serum, and network pharmacology identified 369 potential QFZTP targets for RA, enriched mainly in TNF signaling, PI3K/AKT, MAPK, and apoptosis pathways. Core components (crocetin, epigallocatechin, bergapten) showed stable binding (<-5.0 kcal/mol) to AKT1, EGFR, and PI3K, occupying similar active pockets as native inhibitors. In vitro, QFZTP-containing serum significantly reduced IL-1β-induced MH7A cell viability; inhibited IL-6/TNF-α secretion (comparable to native QFZTP solution); attenuated migration/proliferation; induced apoptosis; downregulated EGFR, p-PI3K, p-AKT, p-p38, p-NF-κB, and Bcl-2; and up-regulated Bax. CONCLUSION: QFZTP synergistically regulates the PI3K/AKT, NF-κB, and MAPK pathways to inhibit synovial cell proliferation and migration, induce apoptosis, and attenuate inflammation. This study provides a robust theoretical and experimental basis for its clinical application in RA.