| Objective: The therapeutic effect of Saussurea involucrata injection on Rheumatoid arthritis rats and the effect on dendritic cell maturation were observed,and the Bone marrow derived dendritic cells were sequenced based on the transcriptome to explore the mechanism of SII in treating RA by inhibiting the related targets and pathways of DC maturation.Methods: A collagen combined with Freund’s complete adjuvant was used to induce a rat model of RA.The rats were divided into control,model,methotrexate(MTX),SII low-dose and SII high-dose groups and given the corresponding treatments.Determination of body weight,ankle circumference,paw thickness and arthritis score in each group of rats;HE staining to detect hyperplasia and inflammatory infiltration in rat ankle synovial tissue.The maturation of DCs in the synovial tissues of the ankle joint and the spleen was detected by immunofluorescence double staining and flow cytometry respectively.ELISA and flow cytometry for Bone marrow derived dendritic cells(BMDC)supernatant inflammatory factor levels and BMDC maturation.Transcriptome sequencing to detect the effect of SII on BMDC maturation.Results: Compared with the Control group,ankle perimeter,thickness of paw and arthritis index,serum pro-inflammatory factor levels were elevated in the Model group,body weight and serum anti-inflammatory factor levels were decreased.HE staining of the ankle joint showed synovial hyperplasia and inflammatory cell infiltration with elevated histopathological score.Immunofluorescence and flow cytometry of the synovial tissue of the ankle joint showed DC maturation in the synovial tissue and spleen of the ankle joint.Compared with the Model group,the SII treatment group could significantly improve these conditions compared to the Model group.In vitro,SII intervention inhibited lipopolysaccharide-induced maturation of BMDC and modulated the expression of inflammatory factors in BMDC supernatants.The transcriptome results showed that the753 co-expressed genes were enriched in the TLR receptor signaling pathway,TNF signaling pathway,chemokine signaling pathways,cytokine-cytokine receptor interaction pathways and other RA-related pathways by GO functional enrichment analysis and KEGG pathway enrichment analysis,with chemokines and cytokines appearing in high frequency in these pathways.Conclusion: SII has a therapeutic effect on RA rats by inhibiting DC maturation,and SII may inhibit DC maturation through the regulation of chemokines,cytokine-mediated chemokine signaling pathways,cytokine-cytokine receptor interaction pathways and other pathways. |
