目的 探讨miRNA-21在支气管哮喘患儿外周血淋巴细胞及支气管哮喘小鼠模型肺组织中的表达,观察其靶基因PTEN在哮喘小鼠模型肺组织中的表达变化。方法 应用real-time PCR方法检测支气管哮喘患儿与正常儿童外周血淋巴细胞miRNA-21的表达,卵蛋白致敏方法建立支气管哮喘小鼠模型,肺泡灌洗液(BALF)细胞计数,HE染色观察气道炎症。应用real-time PCR法检测哮喘组小鼠与对照组小鼠肺组织miRNA-21及其靶基因PTEN的表达,Western blot方法检测哮喘小鼠与对照组小鼠肺组织PTEN蛋白的表达。结果 支气管哮喘患儿外周血淋巴细胞miRNA-21表达水平显著高于正常儿童(P〈0.01)。哮喘小鼠BALF中细胞总数及嗜酸性粒细胞数及肺组织炎症细胞浸润显著高于正常对照组,哮喘组小鼠肺组织miRNA-21mRNA表达水平显著高于正常对照,PTEN mRNA与蛋白水平明显低于正常对照组(P〈0.01)。结论 miRNA-21在支气管哮喘患儿及支气管哮喘小鼠模型中高表达,表明其可能参与支气管哮喘的发病机制。
Objective To investigate the expression of miRNA-21 in peripheral blood lymphocytes of allergic asthma children and lung of asthmatic mice, and its target gene PENT expression in lung of asthmatic mice. Methods The expression of miRNA-21 in peripheral blood lymphocytes of allergic asthma children and normal children was detected by real-time PCR. The asthmatic mice model was established by ovalbumin sensitization. The number of cells in the bronchoalveolar lavage fluid(BALF) of mice in asthmatic group was accessed, and airway inflammation was assessed by HE staining. The expressions of miRNA-21 and its target gene PTEN in lung were detected by real-time PCR. The expression of PTEN protein in lung of mice was detected by Western blot. Results The expression level of miRNA-21 in peripheral blood lymphocytes was increased significantly in allergic asthma children than in normal children(P〈0.01). Compared with control group, The total inflammatory cells or eosinophils in the BALF of mice and inflammatory cells infiltration were increased significantly in asthmatic group than in normal control(P〈0.01). The expression level of miRNA-21 mRNA in lung was higher in asthmatic group than in control group, but lower for PTEN mRNA and protein (P〈0.01). Conclusion The up-regulation expression of miRNA-21might be involved in the pathogenesis of asthma.