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

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

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)比例和乂头框蛋白3(Eoxp3)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:医院皮肤科收集20例过敏性紫癜住院患花和20例健康人.两组间性别、年龄差异无统计学意义(均/>> 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SPSS 16.0软件分析数据.两独M样本均数比较采用,检验.单因素直线相关分析评估各指标的相关性结果过敏件紫瘢组CD4, T淋巴细胞中Foxp3 mRNA表达水平(0.38()± 0.226)显著低于健康对照组"=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%) J = 2.552./J< 0.05,F<>xp3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),￠= 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./>< 0.05),和临床病情评分呈止相关性(r = 0.486.P < 0.05)肾损害组患昔F<xp3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,这町能与过敏性紫癜发病有关,并影响病情进展和预后.

Objective To detrmiine the proportion of CD4 + CD25 + regulator}^ T (Trrg) cells, mRN A expression of the forkhrad box protein 3 ( Foxp3 ) gene, and DNA nirthvlatioii status of the Foxp3 promoter in peripheral (:I)4* T cells from patients with Henoch-Schonlein purpura. Methods Potally. 20 inpatients with Henoch - Schonlein purpura and 20 healthy controls were enrol led from I)('partment of Dermatology, the Second Xiangya Hospital of Central South University between 2() 15 and 2016, and there were no significant differences in the gender and age between the two groups ( both P > 0.05 ).(J)4' 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 SPSS 16.0 software by using two - sample / lest for the comparison between the two groups, and linear correlation analysis for evaluaiing the correlations of the DNA methylation slcitus of the Foxp3 promoter with clinical severity scores and the proportion of CD4 CD254 Treg cells. Results Compared with the healthy control group, the Heiioch-Schonlein purpura group showed significantly (Ircreiisd mRNA expression of the Foxp3 gene in CD4+ T cells (0.380 ± 0.226 rs. 1./= 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 F()xp3 promoter (().712 ± 0.164 vs. 0.453 ± 0.147. t = 3.610. P < 0.01). In the Henoch - Schonlein purpura group, the DNA methylation status of the Foxp3 promoter was negatively correlated with the percentage of CI)4*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 - Schiinlein 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-Schonlein purpura, and affect disease development and prognosis.