中华皮肤科杂志 ›› 2012, Vol. 45 ›› Issue (7): 481-484.

• 论著 • 上一篇    下一篇

特应性皮炎患者外周血调节性T细胞与Th17细胞平衡状态分析

马蕾1,薛海波2,3,周荣佼2,3,安荣贞2,3   

  1. 1. 滨州医学院附属医院皮肤科
    2.
    3. 滨州医学院附属医院
  • 收稿日期:2011-08-17 修回日期:2012-02-23 出版日期:2012-07-15 发布日期:2012-07-02
  • 通讯作者: 马蕾 E-mail:iwf50@sina.com
  • 基金资助:

    山东省自然科学基金;山东省高等学校科技计划项目;滨州医学院科研启动基金

Evaluation of balance between regulatory T cells and T helper 17 cells in patients with atopic dermatitis

  • Received:2011-08-17 Revised:2012-02-23 Online:2012-07-15 Published:2012-07-02
  • Contact: Lei Ma E-mail:iwf50@sina.com

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摘要：

目的 探讨调节性T细胞（Treg 细胞）与Th17细胞失平衡在特应性皮炎（AD）发病机制中的作用。方法 采用流式细胞仪检测41例AD患者外周血Treg细胞与Th17细胞的比例，实时定量逆转录聚合酶链反应检测Treg细胞与Th17细胞转录因子Foxp3、RORγt mRNA的表达水平，酶联免疫吸附试验检测血清中转化生长因子β（TGF-β）、白介素17（IL-17）和白介素23（IL-23）的含量；同时以38例性别、年龄匹配的健康儿童作为对照组。采用独立样本t检验和线性相关进行统计学分析。结果 AD患者组外周血Treg细胞（CD4+CD25+Foxp3+T细胞）比例为2.01% ± 0.57%，Foxp3 mRNA的表达量为0.65 ± 0.19；健康对照组分别为5.04% ± 1.44%和1.71 ± 0.69，AD患者组均明显低于健康对照组（两组比较，t值分别为12.47和9.47，P值均 ＜ 0.01）。AD患者组外周血Th17细胞（CD4+IL-17+T细胞）比例为1.77% ± 0.55%，RORγt mRNA表达量为5.97 ± 1.45；健康对照组分别为0.39% ± 0.15%和1.49 ± 0.57，AD患者组均显著高于健康对照组（两组比较，t值分别为14.82和17.78，P值均＜ 0.01。AD患者组Treg/Th17细胞比值（1.26 ± 0.61）较健康对照组（14.53 ± 5.77）明显降低（t = 14.11，P ＜ 0.01）。急性期AD患者Treg/Th17细胞比值为0.88 ± 0.04，与亚急性期（1.29 ± 0.11，t = 4.02，P ＜ 0.01）和慢性期（2.05 ± 0.24，t = 4.83，P ＜ 0.01）AD患者比较，差异均有统计学意义；AD患者亚急性期与慢性期比较，差异亦有统计学意义（t = 2.89，P ＜ 0.05）。AD患者血清中IL-17和IL-23含量均高于健康对照组（P值均 ＜ 0.01），而TGF-β含量比较，差异无统计学意义（t = 1.96，P ＞ 0.05）。中、重度AD患者疾病严重程度评分与外周血Treg细胞比例呈负相关（r = -0.40，P ＜ 0.05），与Th17细胞比例呈正相关（r = 0.42，P ＜ 0.05）。结论 AD患者外周血Treg和Th17细胞比例、特异性转录因子Foxp3、RORγt mRNA表达水平及血清中相应细胞因子含量发生变化，Treg/Th17细胞失平衡可能参与了AD的发病。

关键词: Th17细胞

Abstract:

Objective To assess the role of imbalance between regulatory T （Treg） cells and T helper 17 （Th17） cells in the pathogenesis of atopic dermatitis（AD）. Methods Peripheral blood was obtained from 41 patients with AD and 38 age- and sex-matched healthy controls. Flow cytometry was performed to determine the percentage of Treg cells （CD4+CD25+Foxp3+ T cells） and Th17 cells（CD4+IL17+ T cells）, real-time quantitative reverse transcription （RT）-PCR to detect the mRNA expressions of Foxp3 and RORγt, which are the specific transcription factors of Treg and Th17 cells respectively. Serum concentrations of transforming growth factor （TGF）-β, IL-17 and IL-23 were measured by enzyme linked immunosorbent assay（ELISA）. Data were statistically assessed by independent-samples t test and Pearson correlation analysis. Results The patients with AD showed an obvious decrease in Treg cell percentage, transcription factor Foxp3 mRNA level and Treg/Th17 ratio （2.01% ± 0.57% vs. 5.04% ± 1.44%, t = 12.47, P ＜ 0.01; 0.65 ± 0.19 vs. 1.71 ± 0.69, t = 9.47, P ＜ 0.01; 1.26 ± 0.61 vs. 14.53 ± 5.77, t = 14.11, P ＜ 0.01）, but a significant increase in peripheral Th17 cell percentage and transcription factor RORγt mRNA level（1.77% ± 0.55% vs. 0.39% ± 0.15%, t = 14.82, P ＜ 0.01; 5.97 ± 1.45 vs. 1.49 ± 0.57, t = 17.78, P ＜ 0.01） compared with the healthy controls. Further comparison revealed that Treg/Th17 ratio was significantly lower in patients with acute AD than in those with subacute AD（0.88 ± 0.04 vs. 1.29 ± 0.11, t = 4.02, P ＜ 0.01） and those with chronic AD（2.05 ± 0.24, t = 4.83，P ＜ 0.01）, statistically different between patients with subacute AD and chronic AD（t = 2.89，P ＜ 0.05）. There was no significant difference in the serum concentration of TGF-β between patients with AD and healthy controls （（15.28 ± 2.34） μg/L vs. （16.56 ± 3.27） μg/L, t = 1.96, P ＞ 0.05）. A significant increase was observed in the serum levels of IL-17 and IL-23 in patients with AD compared with those in the healthy controls（（33.24 ± 7.06） ng/L vs. （11.68 ± 2.67） ng/L, t = 17.96, P ＜ 0.01； （56.35 ± 12.16） ng/L vs. （18.43 ± 3.90） ng/L, t = 18.36, P ＜ 0.01）. In patients with moderate and severe AD, SCORing atopic dermatitis （SCORAD） index was negatively correlated with the percentage of Treg cells （r = －0.40, P ＜ 0.05）, but positively correlated with that of Th17 cells（r = 0.42, P ＜ 0.05）. Conclusions There exists a change in Treg/Th17 ratio, mRNA expressions of RORγt and Foxp3, and serum levels of relevant cytokines in patients with AD, which may lead to immune imbalance and subsequently contribute to the development of AD.

Key words: Th17 cells