Sinomenine inhibits PDGF/PDGFR signaling pathway to reduce RA-FLS migration induced by NETs
This study aims to decipher the mechanism of sinomenine in inhibiting platelet-derived growth factor/platelet-derived growth factor receptor(PDGF/PDGFR)signaling pathway in rheumatoid arthritis-fibroblast-like synoviocyte(RA-FLS)migration induced by neutrophil extracellular traps(NETs).RA-FLS was isolated from the synovial tissue of 3 RA patients and cultured.NETs were extracted from the peripheral venous blood of 4 RA patients and 4 healthy control(HC).RA-FLS was classified into control group,HC-NETs group,RA-NETs group,RA-NETs+sinomenine group and RA-NETs+sinomenine+CP-673451 group.RNA-sequencing(RNA-seq)was conducted to identify the differentially expressed genes between HC-NETs and RA-NETs groups.Sangerbox was used to perform the Gene Ontology(GO)function and the Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment.Cytoscape was employed to build the protein-protein interaction(PPI)network.AutoDock Vina and PyMOL were used for molecular docking of sinomenine with PDGFβ and PDGFRβ.The cell proliferation and migration were determined by the cell counting kit-8(CCK-8)and cell scratch assay,respectively.Western blot was employed to determine the protein level of PDGFRβ.Real-time quantitative polymerase chain reaction(RT-qPCR)was carried out to determine the mRNA levels of matrix metalloproteinases(MMPs).The results revealed that neutrophils in RA patients were more likely to produce NETs.Compared with HC-NETs group,RA-NETs group showed up-regulated expression of PDGFβ and PDGFRβ.Compared with control group,RA-NETs group showed increased cell proliferation and migration and up-regulated protein level of PDGFRβ and mRNA levels of PDGFβ,PDGFRβ,MMP1,MMP3,and MMP9(P＜0.05).Compared with RA-NETs group,RA-NETs+sinomenine group presented decreased cell proliferation and migration and down-regulated protein and mRNA level of PDGFRβ and mRNA levels of MMP1,MMP3,and MMP9(P＜0.05).Compared with RA-NETs+sinomenine group,the proliferation ability of RA-NETs+sinomenine+CP-673451 group decreased(P＜0.05).The findings prove that sinomenine reduces the RA-NETs-induced RA-FLS migration by inhibiting PDGF/PDGFR signaling pathway,thus mitigating RA.
万方数据
维普
