microRNA-33a靶向pendrin调控支气管哮喘气道黏液高分泌-现代医学

microRNA-33a靶向pendrin调控支气管哮喘气道黏液高分泌

单位：1. 榆林市儿童医院儿内二科, 陕西榆林 719000;
2. 榆林市星元医院儿科, 陕西榆林 719000

分类号：R725.6

出版年·卷·期（页码）：2017·36·第三期（420-426）

摘要：

目的：探讨microRNA-33a（miR-33a）在哮喘患儿及哮喘小鼠中的表达及其对气道黏液高分泌的作用。方法：RT-PCR检测哮喘患儿和健康儿童诱导痰，以及卵清蛋白（ovalbumin，OVA）诱导的哮喘小鼠和正常小鼠肺组织中miR-33a的表达；分别鼻滴miR-33a模拟物（miR-33a mimics）及其对照（mimics NC）至哮喘小鼠，ELISA检测各组小鼠支气管肺泡灌洗液（BALF）上清中IL-13、TNF-α及黏蛋白MUC5AC的表达，RT-PCR和western blot检测小鼠肺组织MUC5AC和pendrin（SLC26A4）的表达；双荧光素酶报告法验证miRNA-33a的靶基因。结果：哮喘患儿诱导痰及哮喘小鼠BALF中miR-33a表达显著降低（P<0.05）；哮喘小鼠BALF中嗜酸性粒细胞（Eosnophils，EOS）数目显著增加，IL-13、TNF-α和MUC5AC表达显著升高（P<0.05）；miR-33a mimics滴入导致EOS数目减少，IL-13、TNF-α和MUC5AC表达显著降低（P<0.05），同时小鼠肺组织MUC5AC和pendrin显著下调（P<0.05）；miR-33a mimics明显抑制pendrin（SLC26A4）野生型报告基因的荧光素酶活性（P<0.05）；pendrin（SLC26A4）过表达显著上调miR-33a mimics转染细胞中MUC5AC的表达（P<0.05）。结论：miR-33a通过靶向pendrin抑制黏蛋白MUC5AC的表达，抑制支气管哮喘气道黏液高分泌。

Objective: To investigate the expression of MicroRNA-33a(miR-33a) in bronchial asthma children and murine asthma, and the effect and mechanism of miR-33a on airway mucus hypersecretion. Methods: RT-PCR was employed to determine the differencial expression of miR-33a in induced sputum from bronchial asthma children and healthy individuals, and in lung tissue from ovalbumin(OVA)-induced mouse asthma model and control mice. Asthmatic mice were treated with miR-33a mimics or mimics NC by nasal drip, and the expression of IL-13, TNF-α and MUC5AC in bronchoalveolar lavage fluid(BALF) of the mice were determined by ELISA. RT-PCR and western blot were employed to test the expression of MUC5AC and pendrin in the lung tissue of mice. Dual luciferase report assay was performed to identify the target gene of miR-33a. Results: miR-33a was down-regulated in bronchial asthma children and murine asthma model compared with normal controls(p<0.05). The eosnophils(EOS) numbers, and the expression of IL-13, TNF-α and MUC5AC in BALF from murine asthma group were significantly higher than those of the controls(P<0.05).The EOS numbers, and the expression of IL-13, TNF-α and MUC5AC in BALF from miR-33a mimics group were significantly lower than those of asthma group(P<0.05). The expression of MUC5AC and pendrin in the lung tissue of mice were significantly down-regulated in miR-33a mimics group compared with the asthma group(P<0.05). Transfection of miR-33a mimics significantly suppressed the luciferase activity of wild-type pendrin(SLC26A4)(P<0.05). Overexpression of pendrin(SLC26A4) significantly up-regulated the expression of MUC5AC in cells transfected with miR-33a mimics(P<0.05). Conclusion: miR-33a inhibits the expression of MUC5AC by targeting pendrin, and suppresses bronchial asthma airway mucus hypersecretion.

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