支气管哮喘患儿外周血单核细胞miR-138、RUNX3表达及与Th1/Th2平衡的关系

Expression of miR-138 and RUNX3 in peripheral blood mononuclear cells of children with bronchial asthma and their relationships with Th1/Th2 balance

目的检测支气管哮喘患儿外周血单核细胞miR-138、runt相关转录因子3(RUNX3)表达及Th1/Th2比例,并分析其相关性。方法选取2016年1月-2018年1月本院门诊及病房收治的支气管哮喘患者120例为研究对象,其中急性发作组60例,缓解组60例,选取本院近期同期进行健康体检儿童120例为对照组,利用流式细胞术检测Th1、Th2细胞比例,采用ELISA法检测研究者血清样本中干扰素(IFN)-γ、肿瘤坏死因子(TNF)、白细胞介素(IL)-2、IL-14、IL-10;利用实时定量PCR(qmiR-506)法检测PBMCs中miR-138及RUNX3 mRNA表达情况;利用全自动生化分析仪检测血清IgE水平;利用肺功能测定仪检测第1s用力呼气容积(FEV 1%)。结果与对照组比较,缓解组、急性发作组哮喘患儿FEV 1、RUNX3 mRNA表达水平、Th1细胞亚群比例、Th1/Th2比例、血清IFN-γ、IL-12水平显著降低(P<0.05),血清IgE、外周血单核细胞miR-138表达水平、Th2细胞亚群比例、IL-14、IL-10水平显著升高(P<0.05);与缓解组比较,急性发作组哮喘组患儿FEV 1、RUNX3 mRNA表达水平、Th1细胞亚群比例、Th1/Th2比例、血清IFN-γ、IL-12水平显著降低(P<0.05),血清IgE、外周血单核细胞miR-138表达水平、Th2细胞亚群比例、IL-14、IL-110水平显著升高(P<0.05);支气管哮喘患儿外周血单核细胞miR-138表达水平与RUNX3 mRNA表达水平、Th1/Th2细胞比例负相关(P均<0.05),RUNX3 mRNA表达水平与Th1/Th2细胞比例正相关(P<0.05)。结论哮喘患儿外周血单核细胞miR-138高表达,RUNX3 mRNA低表达,Th1/Th2比例显著降低,二者可能通过调节Th1/Th2细胞比例、介导炎症反应,参与支气管哮喘疾病发生。

Objective To detect the expression of miR-138 and runt-related transcription factor 3(RUNX3)in peripheral blood mononuclear cells of children with bronchial asthma and to analyze their correlation.Methods 120 patients with bronchial asthma admitted to the outpatient and ward of our hospital from January 2016 to January 2018 were selected as the research objects,including 60 cases in the acute attack group and 60 cases in the remission group.120 children who underwent health examination in our hospital during the same period in recent years were selected as the control group.The proportions of Th1 and Th2 cells were detected by flow cytometry,and the levels of serum IFN-γ,TNF,IL-2,IL-4 and IL-10 were detected by ELISA.Real-time quantitative PCR(qRT-PCR)was used to detect the expressions of miR-138 and RUNX3 in PBMCs,the levels of serum IgE was detected by automatic biochemical analyzer,and the forced expiratory volume(FEV 1%)at the 1st second was measured by pulmonary function analyzer.Results Compared with the control group,the expression levels of FEV 1,RUNX3 mRNAs,the ratio of Th1 cell subsets,Th1/Th2 value,and the levels of serum IFN-γand IL-2 of asthmatic children in the remission group and the acute attack group were significantly lower(P<0.05),while the levels of serum IgE,peripheral blood mononuclear cell miR-138,the ratio of Th1 cell subsets,and the levels of IL-4 and IL-10 were significantly higher(P<0.05).Compared with the remission group,the expression levels of FEV 1,RUNX3 mRNAs,the ratio of Th1 cell subsets,Th1/Th2 value,and the levels of serum IFN-γand IL-2 of asthmatic children in the acute attack group were significantly lower(P<0.05),while the levels of serum IgE,peripheral blood mononuclear cell miR-138,the ratio of Th1 cell subsets,and the levels of IL-4 and IL-10 were significantly higher(P<0.05).The expression level of miR-138 in peripheral blood mononuclear cells of children with bronchial asthma was negatively correlated with the expression level of RUNX3 mRNA and the ratio of Th1/Th2 cells(P<0.05),while the expression level of RUNX3 mRNA was positively correlated with the ratio of Th1/Th2 cells(P<0.05).Conclusion The expression of miR-138 in peripheral blood mononuclear cells of asthmatic children is high,the expression of RUNX3 mRNA is low,and the Th1/Th2 is significantly reduced.Both of them may participate in the occurrence of bronchial asthma by regulating the ratio of Th1/Th2 cells and mediating inflammatory reaction.