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探讨(5R)-5-羟基雷公藤内酯醇(雷藤舒)体外在类风湿关节炎(RA)中的免疫调节机制。①CCK-8法检测24h、48h、72h等3个时间段不同浓度雷藤舒对小鼠L929成纤维细胞毒性的影响;②通过3 H胸腺嘧啶脱氧核苷(3 H-TdR)掺入实验,研究雷藤舒对RA患者外周血及滑液中单个核细胞增殖的影响;③流式细胞术检测雷藤舒对RA患者外周血和滑液中Th1细胞、Th17细胞增殖的影响;④ELISA法检测雷藤舒对RA患者外周血及滑液中单个核细胞分泌IFN-γ、TNF-α、IL-17的影响;⑤MTT法研究雷藤舒对RA患者滑膜成纤维细胞(FLS)增殖的影响。结果①雷藤舒≤100nmol/L时在3个时间段(24h、48h、72h)对L929无细胞毒性(P>0.05);②雷藤舒抑制RA患者外周血和滑液中单个核细胞增殖(P<0.05);③雷藤舒抑制Th1(P<0.05)细胞增殖,对Th17细胞增殖有抑制趋势但无统计学意义(P>0.05);④雷藤舒抑制RA患者外周血和滑液中单个核细胞分泌TNF-α、IFN-γ、IL-17水平;⑤雷藤舒抑制RA患者FLS增殖(P<0.05)。雷藤舒对L929细胞无毒性,通过抑制RA患者单个核细胞、Th1、Th17增殖,抑制外周血和滑液中PBMC和SFMC分泌TNF-α、IFN-γ、IL-17,抑制FLS增殖,进而调节RA免疫反应。
To investigate the immunoregulatory mechanism of (5R) -5-hydroxy triptolide (Rheumatoidin) in rheumatoid arthritis (RA) in vitro. (1) The effect of different doses of Thientetra on the toxicity of L929 fibroblasts in mice was detected by CCK-8 assay at 24h, 48h, 72h and so on; (3) By 3 H-TdR incorporation assay, To investigate the effect of Leitengshu on the proliferation of mononuclear cells in peripheral blood and synovial fluid in patients with RA; (3) flow cytometry to detect the effect of Thioushu on the proliferation of Th1 cells and Th17 cells in peripheral blood and synovial fluid of RA patients; (4) The effect of Leitengshu on the secretion of IFN-γ, TNF-α and IL-17 from mononuclear cells in peripheral blood and synovial fluid of patients with rheumatoid arthritis; ⑤MTT method was used to study the effect of Lentucan on the proliferation of synovial fibroblasts (FLS) . Results ① Rattan was not cytotoxic to L929 in three time periods (24h, 48h, 72h) when LeiTuShu ≤100nmol / L (2) LeiTenShu inhibited the proliferation of mononuclear cells in peripheral blood and synovial fluid of RA patients (P0.05); ③ Rattan Shu inhibited the proliferation of Th1 (P <0.05) cells and inhibited the proliferation of Th17 cells but had no statistical significance (P> 0.05). ④ Leishu Shu inhibited the proliferation of peripheral blood and synovial fluid TNF-α, IFN-γ and IL-17 were secreted by mononuclear cells in a dose-dependent manner. ⑤ Rattan Shu inhibited FLS proliferation in RA patients (P <0.05). Lentucan is non-toxic to L929 cells and inhibits the proliferation of mononuclear cells, Th1 and Th17 in RA patients, and inhibits the secretion of TNF-α, IFN-γ and IL-17 from PBMC and SFMC in peripheral blood and synovial fluid. Regulates RA immune response.