桥本甲状腺炎患者外周血调节性T细胞/Th17细胞平衡动态变化的研究

The dynamic changes of Treg/Th17 cells imbalance in patients with Hashimoto thyroiditis

目的探讨桥本甲状腺炎(HT)患者外周血调节性T细胞(Treg)与Th17细胞的动态变化及其意义。方法流式细胞术检测36例HT患者[HT组,根据甲状腺功能分为A(18例)、B(10例)、C(8例)3个亚组]和30例健康对照者外周血单个核细胞(PBMC)中Treg和Th17细胞的比例,实时定量RT-PCR检测Treg和Th17细胞特异性转录因子Foxp3、RORγt mRNA的表达水平,ELISA检测血清中相关细胞因子TGF-β和IL-17A的含量。结果 HT患者PBMC中Treg细胞比例及Foxp3 mRNA表达水平均明显低于健康对照者(t=-10.62、-7.624,P均<0.01),HT亚组间差异具有统计学意义(F=4.414、4.512,P均<0.05);Th17细胞比例、RORγtmRNA表达水平及Th17/Treg比值均明显高于健康对照者(t=11.32、19.25、8.386,P均<0.01),HT亚组间差异均具有统计学意义(F分别为34.947、14.551、24.338,P均<0.01),亚组间两两比较差异均有统计学意义(P均<0.05)。HT患者血清TGF-β的含量较健康对照者略低,但差异无统计学意义(P>0.05);IL-17A的含量均较健康对照者显著升高(P<0.01),HT患者组内各亚组间血清IL-17A的含量差异具有统计学意义(F=11.077,P<0.01),两两比较差异亦具有统计学意义(P均<0.05)。HT患者Th17/Treg比值与血清甲状腺过氧化物酶抗体(TPOAb)、甲状腺球蛋白抗体(TgAb)滴度均呈显著正相关(r=0.349、0.502,P=0.037、0.002)。结论在HT发病过程中,Th17/Treg细胞轴呈动态改变,并且与甲状腺自身抗体滴度正相关,说明Th17/Treg细胞亚群的免疫失平衡可能参与了HT的发生发展。

Objective To investigate the dynamic changes and significance of regulatory T cells（Treg）and Thl7 cells in patients with Hashimoto thyroiditis （HT）. Methods 36 HT patients [ 18 cases with euthyroidism （subset A）, 10 cases with subclinical hypothyroidism（ subset B） ,8 cases with overt hypothyroidism（ subset C）] and 30 healthy controls（NC）were selected. Flow cytometry was performed to detect the percentages of Treg and Thl7 cells. Real-time RT-PCR was employed to analyze the expressions of Foxp3 and RORγt mRNA. Serum concentrations of TGF-13 and IL-17A were measured by ELISA. Results Treg ceils percentages and Foxp3 mRNA levels decreased obviously in HT patients（ t = - 10. 62, -7. 624 respectively and both P 〈 0. 01 ）, one-way ANOVA test in A, B and C subsets revealed that the differences were significant （ F = 4.414,4. 512 respectively and both P 〈 0.05 ）, and a significant increase in Thl7 cells percentages, ROR~/t mRNA levels and the ratio of Thl7/Treg cells （t = 11.32, 19. 25,8. 386 respectively and all P 〈 O. 01 ）compared with NC, one-way ANOVA test in A, B and C subsets revealed that the differences were significant （ F = 34. 947, 14. 551,24. 338 respectively and all P 〈 0.01 ）, and multiple comparison in A, B and C subsets revealed that the differences were significant （ all P 〈 0. 05 ）. There was no statistical difference in serum concentration of TGF-I3 between HT and NC （P 〉 0. 05 ）, while IL-17A levels increased markedly in HT patients（ P 〈 0. 01 ）. In HT patients, the ratio of Th17/Treg ceils positively correlated with the levels of serum thyroperoixidase antibody （ TPOAb）, thyroglobulin antibody （ TgAb ） （ r = 0. 349,0. 502 and P = 0. 037, 0. 002 respectively）. Conclusions During the process of HT, there were dynamic changes of Thl7/Treg cells, and positively correlated with the thyroid specific autoantibodies,which may lead to immune imbalance and contribute to the development of HT.