哮喘患儿外周血单个核细胞microRNA-206表达及意义

Expression of microRNA-206 in peripheral blood mononuclear cells in asthmatic children

目的探讨哮喘患儿外周血单个核细胞（PBMCs）microRNA-206（miR-206）表达在哮喘发生发展中的作用。方法收集27例哮喘患儿发作期和缓解期,以及25名健康对照儿童的外周血标本,分离PBMCs,采用实时定量PCR（qRT-PCR）方法检测PBMCs中microRNA-206及锌指样转录因子4（KLF4）、白介素-17（IL-17）mRNA的相对表达量。结果miR-206、KLF4 mRNA和IL-17 mRNA水平在哮喘发作期、缓解期患儿以及健康对照组儿童三组间的差异均有统计学意义（F=46.58-72.81,P均=0.000）。哮喘发作期的miR-206水平分别低于缓解期和健康对照组,而KLF4 mRNA和IL-17 mRNA水平均分别高于缓解期和健康对照组,差异有统计学意义（P均〈0.01）;缓解期和健康对照组间miR-206、KLF4 mRNA和IL-17 mRNA水平的差异无统计学意义（P均〉0.05）。哮喘患儿发作期,microRNA-206水平与KLF4和IL-17 mRNA相对表达量均呈负相关（r=–0.66、–0.81,P均〈0.01）,KLF4和IL-17 mRNA相对表达量呈正相关（r=0.70,P〈0.01）。结论哮喘患儿外周血单个核细胞microRNA-206表达水平下降,与疾病的发生发展关系密切。

Objective To explore the role of microRNA-206 in peripheral blood mononuclear cells（PBMC） in the pathogenesis and development of childhood asthma. Methods Twenty-seven asthmatic children and 25 healthy controls were enrolled in the study. Peripheral blood mononuclear cells were isolated in both healthy subjects and asthmatic children in acute attack and remission stages. Total RNAs were extracted from PBMC stimulated by PMA and ionomycin, and then the RNA was reversely transcribed into cDNA. The expressions of microRNA-206 and Kruppel-like factor 4（KLF4） and IL-17 mRNA were detected by real-time quantitative PCR（qRT-PCR） method. Results There was significant difference of microRNA-206 levels among asthmatic children in attack stage and in remission stage and normal controls（F=46.58-72.81, P=0.000）. Through pairwise comparison, the microRNA-206 levels of asthmatic children in attack stage were significantly lower than those in remission stage and normal control groups（P〈0.01）. The KLF4 and IL-17 mRNA levels of asthmatic children in attack stage were significantly higher than those in remission stage and normal control groups（P〈0.01）. There was no significant difference of miR-206, KLF4 and IL-17 mRNA between asthmatic children in remission stage and the healthy controls（P〉0.05）. Furthermore, a negative correlation was found between the expression of miR-206 and KLF4（r=–0.66, P〈0.01） and between the expression of miR-206 and IL-17 mRNA（r=–0.81, P〈0.01） in asthmatic children in attack stage. A positive correlation was also found between KLF4 and IL-17 mRNA in asthmatic children in attack stage（r=0.70, P〈0.01）. Conclusions The expression of miR-206 is decreased in asthmatic children, and miR-206 might be involved in the pathogenesis and development of asthma.