TRAF6基因表达对小鼠胶原诱导性关节炎骨髓来源的树突状细胞成熟影响的研究

发布时间：2018-04-22 11:36

  本文选题：类风湿关节炎 + 树突状细胞　； 参考：《桂林医学院》2017年硕士论文

【摘要】：目的:研究肿瘤坏死因子受体(TNFR)相关因子6(TRAF6)与树突状细胞(dendritic cells,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树突状细胞(immature dendritic cell,iDCs),将iDCs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si RNA转染后的iDCs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iDCs TRAF6 mRNA表达明显低于阴性siRNA转染组及未转染组(P0.05),CD80、CD86和MHC-Ⅱ表达明显低于阴性siRNA转染组及未转染组(P0.05),IL-12和TNF-α表达明显低于阴性siRNA转染组及转染组(P0.05)。结论:干扰TRAF6因子在iDCs中表达可抑制DCs的成熟和促炎症因子的释放,TRAF6在RA免疫耐受中起到重要作用。
[Abstract]:Aim: to study the relationship between tumor necrosis factor receptor (TNFR) related factor 6 (TRAF6) and dendritic cells (DCs) maturation. Methods: the collagen induced arthritis (CIA) mouse model was established and bone marrow cells were extracted from the femur of the mice. Recombinant mouse granulocyte-macrophage colony stimulating factor (rmGM-CSFG) and recombinant mouse interleukin-4 (rmIL-4) were used to induce CIA mouse bone marrow cells to differentiate into immature dendritic cells (immature dendritic cells). IDCs was divided into small interfering RNAs (TRAF6-siRNAs) transfection group. Negative siRNA transfection group and non-transfection group. The expression of TRAF6 gene was detected by real-time fluorescence quantitative PCR after transfection. The iDCs matured by lipopolysaccharide stimulated si RNA transfection, the degree of DCs maturation was detected by flow cytometry, and the pro-inflammatory factor released by DCs was detected by enzyme-linked immunosorbent assay (Elisa). Results the success rate of mouse model was 84%. The expression of iDCs TRAF6 mRNA in the TRAF6-siRNA group was significantly lower than that in the negative siRNA transfection group and the untransfected siRNA group. The expression of CD86 and MHC- 鈪，

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