Effect of triptolide on mitochondrial autophagy,activation of NLRP3 inflammatome and pyroptosis of fibroblast synoviocytes in rheumatoid arthritis
Objective To investigate the effects of triptolide on mitochondrial autophagy,NLRP3 inflammatome activation and pyroptosis of fibroblast synovial cells(FLSs)in rheumatoid arthritis.Methods FLSs cells and rheumatoid arthritis FLSs cells were cultured by passage.Rheumatoid arthritis FLSs cells were treated with 0,10,20,40 ng/mL triptolide.The protein expressions of mitochondrial autophagy related proteins LC3B,p62,PHB2,and NLRP3 were detected by Western blotting.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of inflammatory cytokines IL-1β and IL-18.Pyroptosis was detected by flow cytometry,and the expressions of pyroptosis related proteins GSDMD and GSDMD-N were detected by Western blotting.Results After treatment with 10,20,40 ng/mL triptolide,the expression of mitochondrial autophagy associated protein LC3B in FLSs cells of rheumatoid arthritis was significantly reduced in each dose group,and the expression of p62 and PHB2 was upregulated.Expression levels of NLRP3,IL-1β,and IL-18 were significantly decreased.Pyrodeath rate was significantly reduced.Expression of GSDMD protein was significantly increased,and the expression of GSDMD-N protein was decreased in a dose-dependent manner(P<0.05,0.01,0.001).Conclusion Triptolide can inhibit cellular inflammation by inhibiting mitochondrial autophagy,activation of NLRP3 inflammatome and pyroptosis.
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