【摘要】 目的:探讨小鼠腹腔巨噬细胞内DNA-PKcs协同自身免疫调节因子调控Toll样受体(TLRs)的表达水平,阐明外周免疫系统中自身免疫调节因子的调控作用及其意义。方法:小鼠腹腔巨噬细胞分为pEGFPC1/mAire转染组、pEGFPC1/mAire与negative control siRNA共转染组、pEGFPC1/mAire与DNA-PKcssiRNA共转染组、pEGFPC1转染组、pEGFPC1与negative control siRNA共转染组和pEGFPC1与DNA-PKcssiRNA共转染组,采用RT-PCR方法检测各组小鼠腹腔巨噬细胞内TLR1～9的表达水平;采用脂质体转染重组质粒pEGFPC1/mAire和空载质粒pEGFPC1至小鼠腹腔巨噬细胞,采用RT-qPCR方法检测2组细胞TLR1～9的表达水平;采用RT-qPCR方法检测2组转染细胞沉默DNA-PKcs前后TLRs的表达水平。结果:小鼠腹腔巨噬细胞能表达TLR1～9;与转染pEGFPC1/mAire组细胞比较,转染自身免疫调节因子后巨噬细胞TLR1、3和8的表达水平增加(P<0.05),其他组TLRs表达水平无明显变化(P>0.05);DNA-PKcs沉默后,转染pEGFPC1/mAire组细胞TLR1、3和8的表达水平较未沉默组明显下降(P<0.05),而在转染pEGFPC1的细胞内DNA-PKcs沉默前后TLR1-9表达水平均无明显变化(P>0.05)。结论:在小鼠腹腔巨噬细胞内,自身免疫调节因子能够调控TLR1、3和8的表达,其机制可能与DNA-PKcs协同作用有关。更多还原

【Abstract】 Objective To study the expression levels of the Toll like receptor(TLRs) regulated by DNA-PKcs and autoimmune regulator in peritoneal macrophages in mice,and to clarify the function and significance of autoimmune regulator in peripheral immune system.Methods The macrophages were divided into pEGFPC1/mAire transfection group,pEGFPC1/mAire and negative control siRNA co-transfection group,pEGFPC1/mAire and DNA-PKcs siRNA co-transfection group,pEGFPC1 and negative control siRNA co-transfection group,and pEGFPC1 and DNA-PKcs siRNA co-transfection group;the expression levels of TLR1-9 in peritoneal macrophages of the mice in various groups were detected by RT-PCR.The expression levels of TLR1-9 in peritoneal macrophages were detected by RT-qPCR after transfected with pEGFPC1/mAire and pEGFPC1 into the peritoneal macrophages in mice;the expression levels of TLRs in the transfected cells before and after silencing DNA-PKcs were detected by RT-qPCR.Results The peritoneal macrophages in mice could express TLR1-9.Compared with pEGFPC1/mAire transfection group,the expression levels of TLR1,3 and 8 were increased in peritoneal macrophages after transfected with autoimmune regulator(P<0.05) and those in the other groups did not changed significantly(P>0.05).After silencing DNA-PKcs,the expression levels of TLR1,3,and 8 were decreased(P<0.05) in peritoneal macrophages in pEGFPC1/mAire transfection group than thoes in unsilenced group,but the expression levels of TLR1-9 didn’ t have significant changes in macrophages transfected with pEGFPC1 before and after silence(P>0.05).Conclusion Autoimmune regulator can regulate the expression levels of TLR1,3,and 8 in peritoneal macrophages in mice,and the mechnism may be associated with DNA-PKcs interaction effect.更多还原