Notchl信号对银屑病患者外周血Th17细胞分化和功能的影响  

Effect of Notchl signaling pathway on the differentiation and function of Th17 cells in peripheral blood of patients with psoriasis

作　　者：马蕾[1] 薛海波[2] 高梅兰[3] 舒春梅[1] 于娟[1] 张俊花[1] 

机构地区：[1]滨州医学院附属医院皮肤科,山东256603 [2]滨州医学院附属医院内分泌与代谢病科,山东256603 [3]滨州医学院附属医院检验科,山东256603

出　　处：《中华皮肤科杂志》2018年第3期189-193,共5页Chinese Journal of Dermatology

基　　金：山东省医药卫生科技发展计划项目（2016WS0045）

摘　　要：目的 探讨Notch1信号对银屑病患者外周血Th17细胞分化和功能的影响。方法 取35例寻常性银屑病患者与32例健康对照外周血，流式细胞仪检测单个核细胞（PBMC）中Th17细胞占CD4＋ T淋巴细胞的比例，实时荧光定量反转录PCR检测PBMC中维A酸相关孤儿核受体γt（RORγt）、白介素17（IL-17）、Notch1及发状分裂相关增强子1（Hes-1）mRNA表达水平，酶联免疫吸附试验检测血清及培养上清液中IL-17含量。分析Notch1 mRNA的表达与银屑病皮损面积和疾病严重程度评分（PASI）、Th17细胞比例、RORγt mRNA、IL-17 mRNA及蛋白表达水平的相关性。将银屑病患者PBMC分为0（对照组）、2.5、5.0、10.0、20.0 μmol/L γ分泌酶抑制剂（DAPT）处理组，检测DAPT阻断Notch1信号对PBMC中Th17细胞比例、RORγt及IL-17表达水平的影响。结果 银屑病患者PBMC中Th17细胞占CD4＋ T细胞比例为2.863% ± 0.969%，RORγt、Notch1、Hes-1、IL-17 mRNA表达水平分别为5.255 ± 0.998、6.743 ± 1.756、6.384 ± 1.665、6.944 ± 1.626，IL-17血清含量为（36.444 ± 5.936） ng/L，均高于健康对照组［分别为0.604% ± 0.124%、1.530 ± 0.485、1.731 ± 0.456、1.627 ± 0.485、1.698 ± 0.329、（11.762 ± 2.260） ng/L，P 〈 0.01］。银屑病患者Notch1 mRNA表达水平与PASI、Th17细胞比例、RORγt mRNA表达水平、IL-17 mRNA表达水平及血清含量均呈正相关（r值分别为0.584、0.544、0.518、0.549、0.511，均P 〈 0.05）。5种不同浓度DAPT处理组银屑病患者PBMC中Th17细胞比例、RORγt mRNA表达水平、IL-17 mRNA表达水平及培养上清液含量差异有统计学意义（F值分别为79.527、82.239、78.086、80.558，均P 〈 0.01），2.5、5.0、10.0、20.0 μmol/L DAPT组上述指标均低于对照组（均P 〈 0.05），且随着DAPT作用浓度的增加，各指标呈降低趋势。结论 Notch1信号可促进银屑病患者外周血Th17细胞的分化及RORγt 、IL-17、Hes-1的表达。Objective To evaluate the effect of Notch1 signaling pathway on the differentiation and function of Th17 cells in peripheral blood of patients with psoriasis. Methods Peripheral blood samples were obtained from 35 patients with psoriasis and 32 healthy controls. Flow cytometry was performed to determine the proportion of Th17 cells in CD4＋ T cells in peripheral blood mononuclear cells（PBMCs）, real-time RT-PCR to measure the mRNA of retinoic acid receptor-related orphan receptor γt （RORγt）, interleukin （IL）-17, Notch1 and hairy- and-enhancer-of-split-1 （Hes-1）, and enzyme-linked immunosorbent assay （ELISA） to detect levels of IL-17 in the serum and culture supernatant of PBMCs stimulated with phorbol ester, calcium ionophore and brefeldin A. The correlation of Notch1 mRNA with psoriasis area and severity index（PASI） was analyzed, so were its correlation with the proportion of Th17 cells, mRNA of RORγt, and mRNA and protein of IL-17. PBMCs isolated from the patients with psoriasis were divided into 5 groups to be treated with γ-secretase inhibitor N-[N-（3,5-difluorophenacetyl）-l-alanyl]-S-phenylglycine t-butyl ester（DAPT）at different concentrations of 0（control group）, 2.5, 5.0, 10.0 and 20.0 μmol/L, respectively, so as to evaluate the effects of blocking the Notch1 signaling pathway by DAPT on the proportion of Th17 cells in PBMCs, levels of RORγt and IL-17. Results Compared with the healthy controls, patients with psoriasis showed a significant increase in the proportion of Th17 cells in CD4＋ T cells in PBMCs （2.863% ± 0.969% vs. 0.604% ± 0.124%, P 〈 0.01）, mRNA of RORγt （5.255 ± 0.998 vs. 1.530 ± 0.485, P 〈 0.01）, Notch1 （6.743 ± 1.756 vs. 1.731 ± 0.456, P 〈 0.01）, Hes-1 （6.384 ± 1.665 vs. 1.627 ± 0.485, P 〈 0.01） and IL-17 （6.944 ± 1.626 vs. 1.698 ± 0.329, P 〈 0.01）, and serum level of IL-17 （[36.444 ± 5.936] ng/L vs. [11.762 ± 2.260] ng/L, P 〈 0.01）. Among the patients with psoriasis, the mRNA of Notch1 was positivel

关 键 词：银屑病 信号传导 TH17细胞 白细胞介素17 NOTCHL 

分 类 号：R758.63[医药卫生—皮肤病学与性病学]