DNA-PKcs协同自身免疫调节因子调控小鼠腹腔巨噬细胞Toll样受体的表达及其意义

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-PKcs siRNA共转染组、pEGFPC1转染组、pEGFPC1与negative control siRNA共转染组和pEGFPC1与DNA-PKcs siRNA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。
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Expression of Toll like receptor regulated by DNAPKcs and autoimmune regulator in peritoneal macrophages in mice and its significance

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.