【作者】 向瑛；

【导师】 范学工；

【作者基本信息】 中南大学 ， 内科学， 2010， 硕士

【摘要】 目的：探讨体外实验中,抑制性寡脱氧核苷酸(Sup ODN)对小鼠脾脏CD4+T细胞转录因子T-bet、STAT4及STAT6表达的影响。方法：分离小鼠脾脏淋巴细胞,免疫磁珠阳选法纯化CD4+T细胞,流式细胞仪分析分选纯度,在抗-CD3ε抗体、抗-CD28抗体、IL-12作用下,加入Sup ODN或对照寡脱氧核苷酸(Con ODN)培养72小时后,常规提取核蛋白。Western-blot测定核蛋白中磷酸化的磷酸化信号转导子和转录激动子4(Phosphorylationsignal transducers and ctivators of transcription 4,STAT4)、磷酸化信号转导子和转录激动子6 (Phosphorylationsignal transducers and ctivators of transcription 6,STAT6)和T细胞表达的T盒(T box expressed in T cells, T-bet)蛋白表达。结果：与Con ODN组和PBS组相比,Sup ODN组中T-bet和pSTAT4的表达水平明显降低(p<0.05),而pSTAT6表达水平明显升高(p<0.05)。结论：Sup ODN可能通过下调pSTAT4和T-bet的表达,体外抑制小鼠CD4+T细胞向Thl细胞的功能性分化,上调pSTAT6的表达,促进Th2细胞的分化。更多还原

【Abstract】 Objective:To investigate effect of suppressive oligodeoxy-nucleotides (Sup ODN) on expression of T-bet, pSTAT4 and pSTAT6 of CD4+T splenetic lymphocytes in vitro.Methods:The splenetic lymphocytes of BALB/c mice were separated and then CD4+cells were purified with immune magnetic CD4+microbeads(positive selection). The purification was examined by FACS. The co-culture of CD4+cel、anti-CD3ε、anti-CD28、IL-12 and Sup ODN or control oligodeoxynucleotides(Con ODN) was performed for 72 hours. The nucleoprotein was extracted with regular procedure. Expression of pSTAT4(phosphorylated pSTAT4), pSTAT6, T-bet in nucleoprotein were detected by Western-blot.Results:Compared with control ODN and PBS treated group, expression of pSTAT4 and T-bet in Sup ODN treated group decreased significantly (p<0.05), while expression of pSTAT6 in Sup ODN treated group increased significantly (p<0.05).Conclusion:Sup ODN inhibit Thl differentiation of mice CD4+T cell by down regulate expression of pSTAT4 and T-bet, and promote Th2 differentiation by up regulate expression of pSTAT6.更多还原