Abstract

Objective This study assesses the anti-inflammatory and therapeutic effects of Qianghuo Shengshi Tang (QHSST) on rheumatoid arthritis (RA) through network pharmacology and experimental validation and explore the underlying mechanisms. Methods Complete composition analysis of QHSST using UPLC-Q-Orbitrap-MS. And collection potential targets using relevant databases. Protein–protein interaction (PPI) networks were constructed and Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment analyses were conducted. Anti-inflammatory activity of QHSST was evaluated in RAW264.7 cells by assessing apoptosis, NO, interleukin (IL)-6, IL-10, and tumor necrosis factor-α (TNF-α). In rats with wind-cold-dampness paralysis-RA, efficacy of QHSST was confirmed by evaluating foot volumes, arthritis scoring index (AI), blood routine, rheumatoid factor (RF), anti-cyclic citrullinated peptide (CCP) antibody, and hematoxylin and eosin (H&E) staining and inflammatory factors. Enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry were used to examine protein expression in the ankle joints to elucidate the mechanism of QHSST action. Results We identified 44 compounds using UPLC-Q-Orbitrap-MS. Additionally, network pharmacology revealed five active ingredients and seven key targets. GO and KEGG analyses highlighted the involvement of PI3K-Akt signaling and tyrosine kinase inhibitor resistance pathways. Cell studies showed that QHSST reduced apoptosis, NO, IL-6, and TNF-α levels, while increasing IL-10 level in the dexamethasone and 3.125 µg/mL groups, indicating its anti-inflammatory effects. Animal experiments revealed that QHSST and methotrexate significantly decreased AI scores, foot volumes, and blood routine. H&E staining revealed varying degrees of joint harm, accompanied by a decrease in RF, anti-CCP antibody, IL-6, and TNF-α levels, and an increase in IL-10 level, underscoring the efficacy of QHSST in treating RA. ELISA and immunohistochemistry suggest that QHSST may exerts its effects by modulating the EGFR/SRC/PI3 K/AKT/NF-κB pathway. Conclusion Our findings provide a scientific foundation for therapeutic benefits of QHSST in alleviating RA by confirming the anti-inflammatory and RA-relieving properties of the supplement.