IL-34上调Th17细胞的功能促进CIA小鼠TNF-α及IL-6的表达

IL-34 promotes the secretion and expression of TNF-α and IL-6 of CIA by upregulating Th17 cells

目的探讨白细胞介素34(interleukin-34,IL-34)在类风湿关节炎(rheumatoid arthritis,RA)发病中的作用及可能机制。方法重组鼠(recombination mouse,rm)IL-34刺激胶原诱导的关节炎(collagen induced arthritis,CIA)鼠外周血单个核细胞(peripheral blood mononuclear cells,PBMCs),流式细胞检测法(flow cytometry)检测PBMCs中IL-17+CD4+,干扰素(interferon,IFN)γ+CD4+,IL-4+CD4+及IL-10+CD4+T淋巴细胞的百分比;酶联免疫法(enzyme linked immunosorbent assay,ELISA)检测培养液上清中IL-17,IFN-γ,IL-4,IL-10,肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)和IL-6的浓度;实时反转录PCR(real-time reverse transcriptase polymerase chain reaction,RT-PCR)检测PBMCs中TNF-α及IL-6m RNA表达;阻断IL-17后,ELISA法检测培养上清液中TNF-α和IL-6的浓度。结果IL-34刺激CIA鼠PBMCs中Th17细胞的增殖及IL-17的分泌;IL-34刺激CIA鼠PBMCs TNF-α及IL-6m RNA的表达及TNF-a和IL-6的分泌;阻断IL-17后可抑制IL-34促PBMCs分泌TNF-α及IL-6的作用。结论IL-34可通过上调Th17细胞功能刺激TNF-a及IL-6的分泌与表达,参与RA发病,从而为RA的治疗提供新思路和新靶点。

Objective To study the effect of exogenous IL-34 on peripheral blood mononuclear cells（PBMCs）of RA animal model（collagen-induced arthritis,CIA）,explore if IL-34 promotes the production of tumor necrosis factor-α（TNF-α）and IL-6 by Th17 cells.Methods PBMCs of CIA were stimulated by recombination mouse（rm）IL-34,the frequency of IL-17＋CD4＋,interferon（IFN）-γ＋CD4＋,IL-4＋CD4＋and IL-10＋CD4＋T lymphocytes in PBMCs was detected by flow cytometry.The contents of IL-17,IFN-γ,IL-4,IL-10,TNF-αand IL-6 in PBMCs were measured by ELISA following stimulation with anti-CD3/CD28 and rm IL-34.The expressions of TNF-αand IL-6 m RNAs in CIA PBMCs after stimulated with rm IL-34 were detected by RT-PCR;PBMCs were co-cultured with rm IL-34 and anti-IL-17,the secretion of TNF-αand IL-6 were detected by ELISA.Results IL-34 increased the frequency of Th17 cells and stimulated the secretion of IL-17 by PBMCs of CIA.IL-34 increased the secretion of TNF-αand IL-6 and stimulated the expression of TNF-αand IL-6 m RNAs by PBMCs of CIA.Blocking IL-17 by anti-IL-17 antibody decreased the secretion of TNF-αand IL-6 under stimulation with rm IL-34.Conclusion IL-34 promotes the secretion and expression of TNF-αand IL-6 by up-regulated the function of Th17 cells,might be a new treatment target for RA.