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通过体外实验观察雷公藤内酯醇(Triptolide,Tri)对自身反应性T细胞的作用,为以后将Tri用于体内治疗EAE的研究建立基础。采用CCK-8法检测不同浓度Tri对小鼠成纤维细胞株L929生长的影响;采用MOG35-55肽段免疫C57BL/6小鼠建立EAE疾病模型,获取MOG35-55特异性T细胞,3H掺入法检测Tri对其增殖的影响;ELISA方法检测培养上清中IL-17、IFNγ-、TNFα-、IL-4、TGF-β的含量;流式细胞术检测Tri对Th1和Th17细胞分化的影响。结果Tri浓度≤30nmol/L时,对L929细胞生长未见明显细胞毒性;Tri呈剂量依赖性地抑制MOG35-55特异性T细胞增殖,抑制炎症细胞因子IL-17、IFN-γ、TNF-α的表达,并增加IL-4、TGF-β的表达水平,同时抑制na ve T细胞向致病性Th1和Th17细胞的分化。结果表明,Tri可通过抑制MOG35-55特异性T细胞的增殖,改变MOG35-55特异性T细胞分泌细胞因子的格局及抑制Th1和Th17细胞分化的途径影响自身反应性T细胞,从而为Tri体内治疗EAE的研究提供依据,也为临床研究治疗MS药物提供实验基础。
The effect of triptolide (Tri) on autoreactive T cells was observed by in vitro experiments, which laid the foundation for the future study of Tri for the treatment of EAE in vivo. CCK-8 method was used to detect the effect of different concentrations of Tri on the growth of mouse fibroblast cell line L929. The MOG35-55-specific T cells were obtained by immunizing C57BL / 6 mice with MOG35-55 peptide, The effect of Tri on proliferation of Th1 and Th17 cells was detected by flow cytometry (FCM). The levels of IL-17, IFNγ-, TNFα-, IL-4 and TGF- . Results No significant cytotoxicity was observed on the proliferation of L929 cells when Tri concentration was less than 30 nmol / L. Tri inhibited the proliferation of MOG35-55-specific T cells in a dose-dependent manner and inhibited the production of inflammatory cytokines IL-17, IFN-γ, TNF- And increase the expression of IL-4 and TGF-β, meanwhile, inhibit the differentiation of na ve T cells into pathogenic Th1 and Th17 cells. The results showed that Tri could affect autoreactive T cells by inhibiting the proliferation of MOG35-55-specific T cells, altering the pattern of MOG35-55-specific T cells secreting cytokines and inhibiting the differentiation of Th1 and Th17 cells, EAE treatment provides the basis for clinical research for the treatment of MS drugs provide the experimental basis.