Objective The study was to investigate into the relationship between tumor necrosis factor receptor-associated factor 6 (TRAF6) and dendritic cell (DCs) maturation.Methods The bone marrow cells of DBA/1 mice were induced into immature dendritic cells (iDCs) by rmGM-CSF and rmIL-4,and divided into 3 groups (i.e.,the TRAF6 siRNA-transfected group,the control siRNA-transfected group,and the non-transfected group).The iDCs were transfected with siTRAF6,and then stimulated with Lipopolysaccharides (LPS).The mRNA expression of TRAF6 was detected by qRT-PCR.After stimuli by LPS,the expression of CD80,CD86 and MHC-Ⅱ was analyzed by flow cytometry,and the pro-inflammatory factors were detected by ELISA.Results The success rate of building CIA model was 84％.The bone marrow cells were successfully induced into iDCs by co-culture the cytokines rmGM-CSF and rmIL-4.Compared with the control siRNA-transfected group and the non-transfected group,the expression of co-stimulatory molecules in the TRAF6 siRNA-transfected group was decreased by flow cytometry.The pro-inflammatory factors in the TRAF6 siR-NA-transfected group was inferior to the control siRNA-transfected group and the non-transfected group by ELISA.Conclusion The mechanism of DCs maturation might be correlated with TRAF6.%目的 研究肿瘤坏死因子受体相关因子6 (TRAF6)与树突状细胞(DCs)成熟性的关系.方法 建立胶原诱导性关节炎(CIA)小鼠模型,从小鼠股骨中提取骨髓细胞.重组小鼠粒细胞—巨噬细胞集落刺激因子(rmGM-CSF)、重组小鼠白细胞介素-4 (rmIL-4)诱导CIA小鼠骨髓细胞定向分化为未成熟DCs,将未成熟DCs分为针对TRAF6的小干扰RNA (TRAF6-siRNA)转染组、阴性siRNA转染组和未转染组.转染后实时荧光定量PCR检测TRAF6基因表达.脂多糖刺激siRNA转染后的未成熟DCs成熟,流式细胞术检测DCs成熟程度,酶联免疫吸附试验检测DCs释放的促炎症性因子.结果 CIA小鼠造模成功率达84％.转染TRAF6-siRNA组未成熟DCs TRAF6 mRNA表达明显低于阴性siRNA转染组和未转染组,CD80、CD86和MHC-Ⅱ表达明显低于阴性siRNA转染组和未转染组,IL-12和TNF-α表达明显低于阴性siRNA转染组和转染组,均P＜0.05,差异有统计学意义.结论 干扰TRAF6因子在未成熟DCs中表达可抑制DCs的成熟和促炎症因子的释放,TRAF6在类风湿关节炎免疫耐受中起到重要作用.
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