Graves病小鼠模型IL-10水平与Th17细胞功能的检测和意义

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.

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

Objective To study the change of Th17 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 determined by chemiluminescent immunoassay,and serum thyroid stimulating antibodies(TSAb) were determined by cell culture biomethod.The percentages of Th17 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 FT4 levels(r=0.90,P < 0.01).The percentage of Th17 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 expression of IL-17 was increased and IL-10 mRNA expression decreased in the mice GD model.Those results suggest that IL-17 and Il-10 may play an immune regulatory role in the occurrence and development of GD.