SOCS3基因转染对小鼠CD4＋T h细胞分化及炎症细胞因子表达的影响及机制研究

目的：观察SOCS3基因对小鼠CD4＋Th细胞分化及细胞因子表达的影响。方法分离、培养并转染小鼠CD4＋Th细胞，以植物血凝素（PHA）刺激靶细胞，采用逆转录聚合酶链反应（RT‐PCR）检测相应细胞因子基因表达；蛋白质印迹法（West‐ernblot）检测目的细胞因子蛋白表达。结果与对照组相比较，转染组T‐bet、白细胞介素（IL）‐2、干扰素‐γ（IFN‐γ）、信号转导及转录激活因子（STAT）4、IL‐12Rβ2基因表达明显下调，细胞因子信号抑制物（SOCS）3、GATA‐3、IL‐4、IL‐6、IL‐10、STAT6基因表达明显上调，T‐bet、IL‐2、IFN‐γ、STAT4、IL‐12Rβ2蛋白表达明显下调，SOCS3、GATA‐3、IL‐4、IL‐6、IL‐10、STAT6蛋白表达明显上调，差异均有统计学意义（P＜0．01）。结论SOCS3基因转染可上调小鼠CD4＋Th细胞SOCS3基因表达，下调STAT4活化和磷酸化，抑制Th1细胞分化，并下调炎症细胞因子基因和蛋白表达，同时间接促进Th2细胞分化，并上调相应炎症细胞因子基因和蛋白表达。

The effects and mechanism of SOCS3 gene transfection in CD4+Th cell differentiation and expression of inflammatory cytokines of mouse

Objective To investigate the effect and mechanism of adenovirus vector mediated SOCS 3 gene transfection in CD4+ Th cell differentiation and expression of inflammatory cytokines of mouse .Methods The CD4+ Th cells were isolated from spleen of C57bl/6 mouse and cultured .Ad‐SOCS3 were transfected into the CD4+ Th cells .PHA was used for culturing with the CD4+ Th cells .RT‐PCR were used to detect the mRNA expression ,and Western blot were used to detect the protein expression of cytokines .Results Compared with the control group ,the gene and protein expression of T‐bet ,IL‐2 ,IFN‐γ,STAT4 and IL‐12Rβ2 in the transfected group were significantly down‐regulated ,the gene and protein expression of SOCS3 ,GATA‐3 ,IL‐4 ,IL‐6 ,IL‐10 and STAT6 were significantly up‐regulated(P<0 .01) .Conclusion The results indicate that SOCS3 gene transfection can up‐regu‐late SOCS3 mRNA and protein expression in the CD4+ Th cells ,down‐regulate the JAK/STAT pathway ,inhibition of Th1 cell dif‐ferentiation ,and down regulation of inflammatory cytokine gene and protein expression ,and indirectly promote Th2 cell differentia‐tion ,and up the corresponding inflammatory cytokine gene and protein expression .