Treg细胞对帕金森病模型小鼠外周炎症的免疫保护作用

摘要

目的：探讨调节性T细胞(regulatory T cells, Treg cells)对帕金森病(Parkinson’s disease, PD)模型小鼠外周炎症的免疫保护作用。方法：C57BL/6小鼠腹腔注射1-甲基-4-苯基-1,2,3,6-四氢吡啶(1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, MPTP)制备PD模型，1 d后从尾静脉注射磁珠分选并体外激活的Treg细胞，7 d后取小鼠脾脏制备单个核细胞悬液，Trizol法提取细胞总RNA，然后用实时荧光定量聚合酶链式反应法检测小鼠脾脏单个核细胞中促炎因子白细胞介素(interleukin, IL)-17、IL-22和抗炎因子IL-10、转化生长因子-β(transforming growth factor-β, TGF-β)的表达及转录因子维甲酸相关孤核受体-γt(retinoic acid-related orphan receptor-gamma t, ROR-γt)、叉头蛋白P3(forkhead box p3, Foxp3)的表达。结果：PD模型小鼠脾脏单个核细胞中IL-17、IL-22、Foxp3的表达均高于正常小鼠，IL-10、TGF-β、ROR-γt的表达无明显变化；Treg过继转移后，与造模组相比，IL-17、IL-22的表达下降，IL-10、TGF-β的表达升高。结论：Treg细胞过继转移后PD模型小鼠脾脏中致炎因子表达下调，抗炎因子表达上调，提示Treg细胞对PD模型小鼠外周炎症具有免疫保护作用。%Objective: To explore whether regulatory T cells(Treg cells) might play a protective role in peripheral immune inflammation of Parkinson’s disease(PD) model mice. Methods: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP) was intraperitoneally injected into mice to prepare PD model mice. On day 2, Treg cells, which were purified by using magnetic beads and activated in vitro, could be injected into the caudal vein of mice. The method of Trizol and real time quantitative polymerase chain reaction were used to examine the expression levels of pro-inflammatory cytokines, anti-inflammatory cytokines and transcription factors of CD4+T lymphocyte in spleen on day 7 after MPTP injection. Results: Expression levels of the pro-inflammatory cytokines including interleukin IL-17 , IL-22 and transcription factor forkhead box p3(Foxp3), the transcription factor of Treg cells in mononuclear cells of spleen were upregulated. But, the levels of the anti-inflammatory cytokines including IL-10 , transforming growth factor ( TGF )-β and transcription factor of retinoic acid-related orphan receptor-gamma t(ROR-γt), the transcription factor of Th17 cells were not altered in PD mice. However, after adoptive transfer of Treg cells into PD mice, IL-17 and IL-22 were downregulated but IL-10 and TGF-β were raised. Conclusion:Adoptive transfer of Treg cells into PD mice, the expression of pro-inflammatory cytokines were downregulated but the anti-inflammatory cytokines were upregulated, these results suggested that Treg cells might play a protective role in peripheral immune inflammation of PD model mice.