Graves病小鼠模型IL-10水平与Th17细胞功能的检测和意义

Determination of Th17 cellular function and IL-10 in the mice of experimental Graves disease model

目的研究Th17细胞及相关细胞因子IL-17 mRNA与IL-10 mRNA在实验性Graves小鼠体内变化。方法利用TSHR-E(胞外区)的腺病毒免疫小鼠方法建立Graves疾病模型,化学发光法检测血清FT4水平,细胞培养生物学方法检测TSAb活性,流式细胞术(FCM)分析小鼠脾脏细胞中Th17细胞的比例,qRT-PCR检测小鼠脾脏细胞中IL-17和IL-10mRNA水平。结果小鼠血清FT4及TSAb水平升高,TSAb与FT4水平呈显著正相关(r=0.90,P<0.01);Graves小鼠脾细胞中Th17细胞的比例为(2.56±0.43)%,明显高于正常对照组小鼠(1.07±0.27%)%(P<0.05);Graves小鼠脾细胞中IL-17 mRNA的含量明显高于正常对照组(P<0.05),而IL-10 mRNA的含量明显低于正常对照组(P<0.05);且IL-17和IL-10水平变化存在明显的负相关(r=-0.564,P<0.01)。结论 TSHR-E(胞外区)的腺病毒免疫建立的GD模型中,IL-17mRNA增加和IL-10 mRNA则明显下降;提示两者可能在GD的发生及发展中发挥免疫调节作用。

Objective To study the change of Thl7 cells, IL-17 mRNA and IL-10 mRNA in mice of experimental Graves disease （GD）. Methods Mouse model of Grave disease was immunized with Ad-TSHR-E. Serum free T4 concentrations were deter- mined by chemiluminescent immunoassay, and serum thyroid stimulating antibodies （TSAb） were determined by cell culture bio- method. The percentages of Thl7 cells in spleens were determined by flow cytometry. Expression of IL-17 mRNA and IL-10 mRNA in spleen were measured by Real-time RT-PCR. Results Serum free T4 concentrations and anti-TSHR antibodies were increased. There was a positive correlation between serum TSAb values and FF4 levels （r=0.90, P 〈 0.01 ）. The percentage of Thl7 cells in spleens of the GD group （2.56±0.43） % was higher than that in the control group （ 1.07 ±0.27 ） % （P〈0.05）. The level of IL- 10 mRNA expression in spleen from GD mice decreased significantly（P〈0.05）. On the contrary, IL-17 mRNA expression increased markedly （P〈0.05）. There was a negative correlation between amount of IL-10 and IL-17 mRNA（r=-0.564,P〈0.01 ）. Conclusion The ex- pression of IL-17 was increased and IL-10 mRNA expression decreased in the mice GD model. Those results suggest that IL-17 and II-10 may play an immune regulatory role in the occurrence and development of GD.