Human Neutrophil Elastase Induces MUC5AC Overexpression in Chronic Rhinosinusitis Through miR-146a
Human Neutrophil Elastase Induces MUC5AC Overexpression in Chronic Rhinosinusitis Through miR-146a
复制标题
人中性粒细胞弹性蛋白酶通过 miR-146a 诱导慢性鼻窦炎中 MUC5AC 过度表达。
DOI：
10.1177/1945892419871798
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发表时间：
2020-01
影响因子：
2.6
通讯作者：
Luo Qing
中科院分区：
文献类型：
--
作者：
Yan Danqing;Ye Yu;Zhang Jian;Zhao Junmei;Yu Jieqing;Luo Qing
背景慢性鼻窦炎(CRS)的发病机制尚不清楚。MicroRNAs广泛参与多种生理和病理过程，其中microRNA-146a(miR-146a)在先天免疫、炎症反应等病理生理过程中发挥重要作用。粘蛋白(MUCs)是分泌性粘液的重要组成部分，其中MUC5AC是正常呼吸道分泌的主要粘蛋白。目的通过miR-146a检测人中性粒细胞弹性蛋白酶(HNE)诱导CRS细胞MUC5AC的过度表达。方法采用实时定量聚合酶链式反应(qRT-PCR)检测鼻窦黏膜miR-146a、HNE、表皮生长因子受体(EGFR)和MUC5AC的表达。在原代培养的人鼻上皮细胞(HNECs)中检测EGFR、磷酸化EGFR(PEGFR)和MUC5AC的表达。我们通过用miR-146a模拟物和阴性对照(NC)转染HNEC，检测miR-146a、MUC5AC、EGFR和pEGFR的表达。此外，利用双荧光素酶报告基因分析验证了EGFR是否为hsa-miR-146a靶基因。结果在有无鼻息肉的CRS患者中，miR-146a表达显著下调，HNE、EGFR和MUC5AC表达上调。在体外细胞实验中，使用EGFR特异性抑制剂(AG1478)后，MUC5AC的表达显著下调。加入miR-146a抑制剂后，miR-146a表达下调，而MUC5AC表达上调。MiR-146a可抑制正常原代HNEC的MUC5AC表达，下调pEGFR的表达。双荧光素酶报告基因检测结果显示，与pGL3+miR-146a模拟组相比，pGL3-EGFR-3‘UTR+miR-146a模拟组的荧光素酶活性受到明显抑制，提示EGFR是miR-146a的靶基因。结论在HNE诱导的CRS中，miR-146a通过抑制EGFR的激活而下调MUC5AC的表达，EGFR是miR-146a的靶基因。
Background The pathogenesis of chronic rhinosinusitis (CRS) is not yet clear. microRNAs are widely involved in a number of physiological and pathological processes, of which microRNA-146a (miR-146a) plays an important role in innate immunity, inflammatory response, and other pathophysiological processes. Mucins (MUCs) are important components of secreted mucus, of which MUC5AC is the major MUC secreted in the normal airway. Objective This study was performed to examine human neutrophil elastase (HNE)-induced MUC5AC overexpression in CRS via miR-146a. Methods miR-146a, HNE, epidermal growth factor receptor (EGFR), and MUC5AC expression in the sinonasal mucosa were determined using quantitative real-time polymerase chain reaction (qRT-PCR). EGFR, phosphorylated EGFR (pEGFR), and MUC5AC expression were determined in primary cultures of human nasal epithelial cells (HNECs). We examined the expression of miR-146a, MUC5AC, EGFR, and pEGFR by transfecting HNECs with miR-146a mimics and negative control (NC). Moreover, dual-luciferase reporter gene assays were used to validate EGFR as an hsa-miR-146a target gene. Results miR-146a was significantly downregulated, and HNE, EGFR, and MUC5AC were upregulated in CRS patients both with and without nasal polyps. In the in vitro cell experiment, MUC5AC was significantly downregulated after use of an EGFR-specific inhibitor (AG1478). Upon addition of miR-146a inhibitor, miR-146a was downregulated, while MUC5AC was upregulated. MUC5AC was suppressed in normal primary HNECs by miR-146a mimic and pEGFR was downregulated. The results of dual-luciferase reporter assays showed that the luciferase activities were markedly inhibited in the pGL3-EGFR-3′ UTR+miR-146a mimic group compared with the pGL3+ miR-146a mimic group, suggesting that EGFR is a target gene for miR-146a. Conclusion In HNE-induced CRS, miR-146a downregulates the expression of MUC5AC by inhibiting the activation of EGFR, and EGFR is a target gene of miR-146a.
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DOI：
10.3904/kjim.2005.20.4.275
发表时间：
2005-12
期刊：
The Korean journal of internal medicine
影响因子：
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作者：
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通讯作者：
Park SH