Th1、Th2、Th17细胞变化及环境因素与习惯性流产的关系研究

Study of the association of Th1, Th2, Th17 cells and environmental factors with habitual abortion

目的 探讨Th1、Th2和Th17细胞的变化、转录因子T-bet、GATA-3和STAT-3 mRNA的表达及环境因素与习惯性流产的关系.方法 选取2011年6月～2012年9月河南省开封市妇产医院就诊的50例习惯性流产非孕期患者作为病例组（HA组）,同期某社区50例正常非孕妇女为健康对照组（NNP组）,分别抽取抗凝静脉血5 mL,应用聚多糖密度梯度离心法分离外周血单个核细胞（PBMC）,采用流式细胞仪检测三种亚群细胞的比例,酶联免疫吸附法（ELISA）检测IFN-γ、IL-4和IL-17的含量,实时荧光定量PCR检测T-bet 、GATA-3和STAT-3 mRNA的表达.同时进行环境因素问卷调查,比较两组各环境因素的差异.结果 ①HA组患者Th1、Th17细胞百分比[（11.66±3.45）％、（3.65±0.75）％]均明显高于NNP组[（7.05±1.83）％、（0.72±0.11）％],差异有统计学意义（P＜0.05）,Th2细胞的百分比[（1.15±0.33）％]低于NNP组[（3.59±0.62）％],差异有统计学意义（P＜0.05）.②HA组IFN-γ和IL-17在的含量分别为（1163.27±45.25）μg/L和（162.26±25.28）μg/L,明显高于NNP组[（736.25±44.32）、（57.45±11.26） μg/L],差异有统计学意义（P＜0.05）,而IL-4的含量[（118.45±26.75） μg/L]低于NNP组[（220.45±24.63）μg/L],差异有统计学意义（P＜0.05）.③与NNP组比较,HA组T-bet和STAT-3 mRNA表达均明显升高,而GATA-3 mRNA的表达明显降低,差异均有统计学意义（均P＜ 0.05）.④HA组BMI≥24.0 kg/m2者所占比例为58％,明显高于NNP组（28％）,差异有统计学意义（P＜0.05）.结论 习惯性流产患者IFN-γ、IL-17的水平较高,而IL-4的水平较低,提示习惯性流产患者体内存在T细胞亚群分化失衡,且此种失衡可能与转录因子T-bet、GATA-3和STAT-3 mRNA的调控有关,同时肥胖可增加习惯性流产发生的可能性.

Objective To explore the association of Th1,Th2,Th17 cells and environmental factors with habitual abortion and the expression of relevant transcription factor T-bet、GATA-3 and STAT-3 mRNA.Methods 50 non-pregnant habitual abortion （HA group） patients from June 2011 to September 2012 were collected,while 50 normal non-pregnancy （NNP group） patients were served as controls.Peripheral blood mononuclear cell （PBMC） was isolated by Ficoll.The proportions of Th1,Th2 and Th17 cells were detected through flow cytometry.IFN-γ, IL-4 and IL-17 levels in the supernatant were detected by enzyme-linked immunosorbent assay （ELISA）.The messenger RNA （mRNA） expression levels of T-bet 、GATA-3 and STAT-3 were amplified by Real-time polymerase chain reaction （RTQ-PCR）.At the meantime,the environmental factors were studied questionnaire survey.Results ① The percentages of Th1 and Th17 cells of patients in HA group [（11.66±3.45）％,（3.65±0.75）％] were significantly higher than those of NNP group [（7.05±1.83）％,（0.72±0.11）％],the differences were statistically significant （P ＜ 0.05）.The percentage of Th2 cells in HA group [（1.15±0.33）％] was lower than that in NNP group [（3.59±0.62）％],the difference was statistically significant （P ＜ 0.05）.② The levels of IFN-γand IL-17 in the HA group were （1163.27±45.25） μg/L and （162.26±25.28） μg/L respectively,significantly higher than those in NNP group [（736.25±44.32）,（57.45±11.26） μg/L],the differences were statistically significant （P ＜ 0.05）.Meanwhile,the level of IL-4 in HA group was greatly lower than that in that in NNP group [（118.45±26.75） μg/L vs （220.45±24.63） μg/L],the difference was statistically significant （P ＜ 0.05）.③ Compared with the NNP group,the level of T-bet and STAT-3 mRNA in the HA group were significantly increased,and the level of GATA-3 mRNA was significantly decreased,the differences were statistically significant （all P ＜ 0.05）.④ The percentages of patients with BMI≥ 24.0 kg/m2 in HA group （58％） was greatly higher than that of NNP group （28％）,the difference was statistically significant （P ＜ 0.05）.Conclusion There is T cells differentiation and imbalance in the habitual abortion patients,accompanying with over-production of IFN-γ and IL-17 in PBMC while lower-production of IL-4.The imbalance differentiation of T cells in habitual abortion patients may be regulated by transcription factor T-bet 、GATA-3 and STAT-3.Higher BMI （≥ 24.0 kg/m2） can also increase the incidence of HA in Chinese women.