ORMDL3 may participate in the pathogenesis of bronchial epithelial-mesenchymal transition in asthmatic mice with airway remodeling
ORMDL3 may participate in the pathogenesis of bronchial epithelial-mesenchymal transition in asthmatic mice with airway remodeling
复制标题
DOI：
10.3892/mmr.2017.7972
复制
发表时间：
2018-01-01
影响因子：
3.4
通讯作者：
Shang, Yunxiao
中科院分区：
文献类型：
--
作者：
Cheng, Qi;Shang, Yunxiao
哮喘是儿童常见的慢性呼吸道疾病，是由遗传和环境因素之间复杂的相互作用引起的。 Orosumucoid-like 3 (ORMDL3) 是一个与哮喘密切相关的候选基因；然而，其根本机制尚不清楚。 ORMDL3 调节金属蛋白酶和转化生长因子的表达，ORMDL3 转基因小鼠表现出气道重塑增强。因此，ORMDL3可能与气道重塑有关。本研究试图检查 ORMDL3 与哮喘小鼠气道重塑严重程度之间的关联，并确定 ORMDL3 是否诱导支气管上皮中的上皮间质转化 (EMT)。为此，BALB/c 小鼠被随机分配到对照组和哮喘组。在卵清蛋白 (OVA) 攻击的第 3、7 和 14 天收集肺组织。然后通过苏木精和曙红以及马森染色观察哮喘小鼠的气道重塑。通过透射电子显微镜评估支气管上皮的形态变化。采用Western blotting和逆转录定量聚合酶链反应（RT-qPCR）对哮喘小鼠气道重塑不同时间点的EMT相关指标E-钙粘蛋白（E-cad）、成纤维细胞特异性蛋白1（FSP1）和波形蛋白（VIM）进行评估，以检测EMT的变化趋势。然后，通过免疫组织化学观察ORMDL3的定位，并分别通过Western blotting和RT-qPCR检测其蛋白和mRNA表达。进一步将表达ORMDL3的质粒转染支气管上皮细胞系16HBE14o-，通过免疫荧光、Western blotting和RT-qPCR检测E-cad、FSP-1和VIM表达的差异；通过显微镜评估细胞侵袭能力。结果显示，支气管上皮中 ORMDL3 的表达与体内气道重塑和 EMT 进展相关。将 ORMDL3 转染至 16HBE 14o 细胞中可在体外诱导 EMT。总而言之，这些发现表明 ORMDL3 可能调节支气管上皮的 EMT，从而影响哮喘的气道重塑。
Asthma is a common chronic respiratory disease in children that is caused by a complex interaction between genetic and environmental factors. Orosomucoid-like 3 (ORMDL3) is a candidate gene that has been strongly associated with asthma; however, the underlying mechanisms are unknown. ORMDL3 regulates the expression of metalloproteinases and transforming growth factor-, and ORMDL3 transgenic mice exhibit increased airway remodeling. Therefore, ORMDL3 may be associated with airway remodeling. The present study attempted to examine the associations between ORMDL3 and the severity of airway remodeling in asthmatic mice, and also to determine whether ORMDL3 induces epithelial-mesenchymal transition (EMT) in the bronchial epithelium. For this purpose, BALB/c mice were randomly assigned to control and asthma groups. Lung tissues were collected on days 3, 7 and 14 of the ovalbumin (OVA) challenge. Airway remodeling in asthmatic mice was then observed by hematoxylin and eosin, and Masson staining. Morphological changes in the bronchial epithelium were assessed by transmission electron microscopy. The EMT-associated indicators E-cadherin (E-cad), fibroblast-specific protein 1 (FSP1) and Vimentin (VIM) were assessed by western blotting and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) at different time points of airway remodeling in asthmatic mice to detect the trend in EMT. Then, the localization of ORMDL3 was observed by immunohistochemistry, and its protein and mRNA expression was examined by western blotting and RT-qPCR, respectively. Furthermore, the bronchial epithelial cell line 16HBE14o-was transfected with an ORMDL3-expressing plasmid, and the differences in E-cad, FSP-1 and VIM expression were detected by immunofluorescence, western blotting and RT-qPCR; the cell invasive ability was assessed by microscopy. The results revealed that ORMDL3 expression in the bronchial epithelium was associated with airway remodeling and EMT progression in vivo. Transfection of ORMDL3 into 16HBE 14o-cells in vitro induced EMT. Taken together, these findings suggest that ORMDL3 may regulate EMT in the bronchial epithelium, thereby affecting airway remodeling in asthma.