中华皮肤科杂志 ›› 2019, Vol. 52 ›› Issue (3): 162-166.doi: 10.3760/cma.j.issn.0412-4030.2019.03.003

• 论著 • 上一篇    下一篇

过敏性紫癜患者CD4+ T细胞叉头框蛋白3基因甲基化水平及其与调节性T细胞的关系

树叶1,罗鸯鸯1,罗勇奇1,汤建萍1,肖嵘2   

  1. 1湖南省儿童医院皮肤科,长沙 410007; 2中南大学湘雅二医院皮肤科,长沙 410008
  • 收稿日期:2018-08-02 修回日期:2018-12-17 出版日期:2019-03-15 发布日期:2019-03-04
  • 通讯作者: 肖嵘 E-mail:xiaorong65@aliyun.com
  • 基金资助:
    湖南省医药卫生科研计划项目(B2016039)

DNA methylation status of the forkhead box protein 3 gene in CD4+ T cells of patients with Henoch-Schönlein purpura and its correlation with regulatory T cells

Shu Ye1, Luo Yangyang1, Luo Yongqi1, Tang Jianping1, Xiao Rong2   

  1. 1Department of Dermatology, Hunan Children′s Hospital, Changsha 410007, China; 2Department of Dermatology, The Second Xiangya Hospital of Central South University, Changsha 410008, China
  • Received:2018-08-02 Revised:2018-12-17 Online:2019-03-15 Published:2019-03-04
  • Contact: Xiao Rong E-mail:xiaorong65@aliyun.com
  • Supported by:
    Medical and Health Research Project of Hunan Province (B2016039)

摘要: 【摘要】 目的 检测过敏性紫癜患者外周血CD4+ T淋巴细胞中CD4+CD25+调节性T细胞(Treg)比例和叉头框蛋白3(Foxp3)基因mRNA表达以及启动子区甲基化水平。方法 2015—2016年在中南大学湘雅二医院皮肤科收集20例过敏性紫癜住院患者和20例健康人,两组间性别、年龄差异无统计学意义(均P > 0.05)。分离受试者外周血CD4+ T淋巴细胞,荧光实时定量PCR试剂盒检测Foxp3基因mRNA,流式细胞仪检测CD4+CD25+Treg比例,亚硫酸氢钠测序技术检测Foxp3基因启动子甲基化水平。采用SPSS16.0软件分析数据,两独立样本均数比较采用t检验,单因素直线相关分析评估各指标的相关性。结果 过敏性紫癜组CD4+ T淋巴细胞中Foxp3 mRNA表达水平(0.380 ± 0.226)显著低于健康对照组,t = 9.503,P < 0.01,CD4+CD25+Treg比例(1.668% ± 0.959%)显著低于对照组(2.741% ± 1.131%),t = 2.552,P < 0.05,Foxp3基因启动子区甲基化水平(0.712 ± 0.164)显著高于对照组(0.453 ± 0.147),t = 3.610,P < 0.01。患者组Foxp3启动子区甲基化水平与CD4+CD25+Treg百分比呈显著负相关(r = -0.490,P < 0.05),和临床病情评分呈正相关性(r=0.486,P < 0.05)。肾损害组患者Foxp3 基因启动子区甲基化水平显著高于无肾损害组(P < 0.05)。结论 过敏性紫癜患者CD4+ T细胞Foxp3基因启动子区甲基化水平升高,下调Foxp3基因表达和CD4+CD25+Treg比例,这可能与过敏性紫癜发病有关,并影响病情进展和预后。

关键词: 紫癜, 过敏性; T淋巴细胞, 调节性; DNA甲基化; 基因, Foxp3

Abstract: 【Abstract】 Objective To determine the proportion of CD4+CD25+ regulatory T (Treg) cells, mRNA expression of the forkhead box protein 3 (Foxp3) gene, and DNA methylation status of the Foxp3 promoter in peripheral CD4+ T cells from patients with Henoch-Sch?nlein purpura. Methods Totally, 20 inpatients with Henoch-Sch?nlein purpura and 20 healthy controls were enrolled from Department of Dermatology, the Second Xiangya Hospital of Central South University between 2015 and 2016, and there were no significant differences in the gender and age between the two groups (both P > 0.05). CD4+ T cells were isolated from the peripheral blood samples of these subjects. Real-time fluorescence-based quantitative PCR was performed to detect the mRNA expression of the Foxp3 gene, flow cytometry to determine the proportion of CD4+CD25+ Treg cells, and sodium bisulfite sequencing PCR (BSP) to determine the DNA methylation status of the Foxp3 promoter. Statistical analysis was carried out with SPSS16.0 software by using two-sample t test for the comparison between the two groups, and linear correlation analysis for evaluating the correlations of the DNA methylation status of the Foxp3 promoter with clinical severity scores and the proportion of CD4+CD25+ Treg cells. Results Compared with the healthy control group, the Henoch-Sch?nlein purpura group showed significantly decreased mRNA expression of the Foxp3 gene in CD4+ T cells (0.380 ± 0.226 vs. 1, t = 9.503, P < 0.01), proportion of CD4+CD25+ Treg cells (1.668% ± 0.959% vs. 2.741% ± 1.131%, t = 2.552, P < 0.05), but significantly increased DNA methylation status of the Foxp3 promoter (0.712 ± 0.164 vs. 0.453 ± 0.147, t = 3.610, P < 0.01). In the Henoch-Sch?nlein purpura group, the DNA methylation status of the Foxp3 promoter was negatively correlated with the percentage of CD4+CD25+ Treg cells (r = -0.490, P < 0.05), but positively correlated with the clinical severity scores (r = 0.486, P < 0.05). The DNA methylation level of the Foxp3 promoter was significantly higher in the patients with renal impairment than in those without renal impairment (P < 0.05). Conclusion The patients with Henoch-Sch?nlein purpura showed increased DNA methylation status of the Foxp3 promoter in CD4+ T cells, decreased mRNA expression of the Foxp3 gene and proportion of CD4+CD25+ Treg cells, which may be related to the occurrence of Henoch-Sch?nlein purpura, and affect disease development and prognosis.

Key words: Purpura, Schoenlein?Henoch, T?lymphocytes, regulatory, DNA methylation, Genes, Foxp3