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目的探讨外源性白三烯B4(LTB4)对胶原诱导型关节炎(collagen-induced arthritis,CIA)小鼠脾细胞调节性T细胞(Treg)和Th17细胞分化的调节,进一步阐明LTB4在类风湿关节炎(RA)发病中的作用机制。方法建立CIA小鼠模型,取造模d28的脾细胞,体外实验分析外源性LTB4对Treg和Th17细胞分化的影响。应用流式细胞术检测CD4+CD25+Foxp3+细胞的数量,荧光定量PCR技术检测调控Treg和Th17细胞分化的特异性转录因子Foxp3和RORγt的mRNA的表达,酶联免疫吸附(ELISA)方法检测培养细胞上清IL-17的含量。结果成功建立CIA小鼠模型;造模d28分离小鼠脾细胞,体外培养加入鸡Ⅱ型胶原(CⅡ)共同孵育,随着LTB4浓度增加(0.01、0.1、1μmol·L-1),CD4+CD25+Foxp3+细胞数量相应减少,Foxp3 mRNA的表达相应降低;相反,IL-17的含量相应增加,RORγt mRNA的表达相应升高。结论LTB4抑制CIA模型脾细胞Treg细胞的分化,促进Th17细胞的分化,提示LTB4在CIA发病过程中具有一定的促炎活性。
Objective To investigate the regulatory effect of exogenous leukotriene B4 (LTB4) on the differentiation of Treg and Th17 cells in collagen-induced arthritis (CIA) mice and to further clarify the role of LTB4 in rheumatoid arthritis The mechanism of action in the pathogenesis of arthritis (RA). Methods The CIA mouse model was established, and the spleen cells of model d28 were obtained. The effect of exogenous LTB4 on the differentiation of Treg and Th17 cells was analyzed in vitro. The number of CD4 + CD25 + Foxp3 + cells was detected by flow cytometry. The mRNA expression of Foxp3 and RORγt, which regulate the differentiation of Treg and Th17 cells, was detected by real-time fluorescence quantitative PCR. The cultured cells were detected by enzyme linked immunosorbent assay (ELISA) The content of supernatant IL-17. Results CIA mouse model was established successfully. Spleen cells were isolated from mice by modeling d28 and cultured in vitro. Collagen II (CⅡ) was used to incubate the cells. With the increase of LTB4 concentration (0.01,0.1,1μmol·L-1), CD4 + CD25 + Foxp3 + cells decreased correspondingly, and the expression of Foxp3 mRNA decreased accordingly. On the contrary, the content of IL-17 increased correspondingly and the expression of RORγt mRNA increased correspondingly. Conclusion LTB4 can inhibit the differentiation of Treg cells and promote the differentiation of Th17 cells in CIA model spleen cells, suggesting that LTB4 may have some proinflammatory activity in the pathogenesis of CIA.