摘要目的 分析斑秃患者外周血Th9细胞及其相关细胞因子的表达与斑秃严重程度、病程等的关系,探讨其在斑秃发病中的意义.方法 2017年5-12月选取就诊于复旦大学附属华山医院皮肤科门诊的74例斑秃患者作为斑秃组,57例本院健康体检者作为对照组.采集受试者外周血,酶联免疫吸附试验(ELISA)检测血清中白细胞介素(IL)-9、IL-4、转化生长因子(TGF)-β1、干扰素(IFN)-γ等细胞因子表达量,流式细胞仪检测外周血单个核细胞(PBMC)中Th9细胞(CD4+IL-9+)所占比例,实时荧光定量PCR法检测PBMC中IL-9及转录因子PU.1 mRNA表达.Excel 2017录入数据,SPSS17.0统计软件分析数据,Graphpad prism 6软件绘制统计图.两组独立样本比较使用t检验,相关分析采用Spearman相关检验.结果 斑秃组血清IL-9水平(190.40±12.33 ng/L)显著低于对照组(288.10±17.38 ng/L,=4.71,P＜ 0.01),TGF-β1(6 191.00±355.50 ng/L)和IFN-γ水平(15.71±3.00 ng/L)均高于对照组(4 026.00±258.00 ng/L、8.79±0.60 ng/L),差异有统计学意义(t=4.41、2.001,均P＜0.05),但两组IL-4水平差异无统计学意义(P＞0.05).重症斑秃患者血清中IFN-γ水平显著低于轻症患者(P=0.02),患者血清中IFN-γ水平与脱发严重度(SALT)分级呈负相关(rs=-0.298,P=0.010);重症与轻症患者血清中IL-9、IL-4及TGF-β1水平差异均无统计学意义(P＞0.05).活动期与稳定期患者、病程＜6个月与＞6个月患者血清中IL-9、IL-4、TGF-β1及IFN-γ水平差异均无统计学意义(P＞0.05).斑秃组PBMC中Th9细胞比例显著低于对照组(t=2.04,P=0.045),且IL-9、PU.1 mRNA相对含量均显著低于对照组(t=2.12、2.178,均P＜0.05).结论 斑秃患者外周血IL-9水平及Th9细胞比例明显减少,Th9细胞及其相关细胞因子可能参与斑秃的发病.

abstractsObjective To investigate the relationship of the expression of peripheral T helper type 9 (Th9) cells and their related cytokines with the severity and clinical course of alopecia areata,and to explore their significance in the occurrence of alopecia areata.Methods From May to December in 2017,74 outpatients with alopecia areata enrolled from Department of Dermatology,Huashan Hospital,Fudan University served as the alopecia areata group,and 57 health checkup examinees in Huashan Hospital served as the control group.Peripheral blood samples were obtained from the patients and controls.Enzyme-linked immunosorbent assay (ELISA) was performed to detect the serum levels of interleukin (IL)-9,IL-4,transforming growth factor (TGF)-β1 and interferon (IFN)-γ,flow cytometry to determine the proportion of Th9 cells (CD4+IL-9+ T helper cells) in peripheral blood mononuclear cells (PBMC),and real-time fluorescence-based quantitative PCR to measure the mRNA expression of IL-9 and PU.1 in the PBMCs.Data were recorded in Microsoft Excel 2017 software,and statistically analyzed with SPSS17.0 software using two-sample t test and Spearman rank correlation analysis.Statistical charts were drawn with Graphpad prism 6 software.Results Compared with the control group,the alopecia areata group showed significantly decreased serum level of IL-9 (190.40 ± 12.33 ng/L vs.288.10 ± 17.38 ng/L,t =4.71,P ＜ 0.01),but significantly increased serum levels of TGF-β1 (6 191.00 ± 355.50 ng/L vs.4 026.00 ± 258.00 ng/L,t =4.41,P ＜ 0.05) and IFN-γ(15.71 ± 3.00 ng/L vs.8.79 ± 0.60 ng/L,t =2.001,P ＜ 0.05).However,there was no significant difference in the serum level of IL-4 between the alopecia areata group and control group (P ＞ 0.05).The serum level of IFN-γ was significantly lower in the patients with severe alopecia areata than in the patients with mild alopecia areata (P =0.02),and the serum level of IFN-γ in the patients with alopecia areata was negatively correlated with the severity of alopecia tool (SALT) score (ru =-0.298,P =0.010).There were no significant differences in the serum levels of IL-9,IL-4 and TGF-β1 between the patients with severe alopecia areata and those with mild alopecia areata (all P ＞ 0.05).The serum levels of IL-9,IL-4,TGF-β1 and IFN-γdid not differ between the patients with active alopecia areata and those with stable alopecia areata,as well as between the patients with clinical course of ＜ 6 months and those with clinical course of ＞ 6 months (P ＞ 0.05).The alopecia areata group showed significantly decreased proportion of Th9 cells in the PBMCs (t =2.04,P =0.045) and mRNA expression of IL-9 and PU.1 (t =2.12,2.178,both P ＜ 0.05) compared with the control group.Condusion The serum level of IL-9 and proportion of peripheral blood Th9 cells both decrease in patients with alopecia areata,and Th9 cells and their related cytokines may be involved in the occurrence of alopecia areata.