The role of autophagy in the overexpression of MUC5AC in patients with chronic rhinosinusitis
The role of autophagy in the overexpression of MUC5AC in patients with chronic rhinosinusitis
复制标题
自噬在慢性鼻-鼻窦炎患者MUC5AC过表达中的作用
DOI：
10.1016/j.intimp.2019.03.028
复制
发表时间：
2019
影响因子：
5.6
通讯作者：
Luo Qing
中科院分区：
文献类型：
--
作者：
Ye Yu;Zhao Junmei;Ye Jing;Jiang Xiaoyue;Liu Haitao;Xie Yanhua;Zhang Jian;Luo Qing
背景自噬是一种保护机体的溶酶体降解途径，对于细胞生存和分化至关重要。粘蛋白（MUCs）是分泌性粘液的重要组成部分，粘蛋白（MUC）5AC是正常气道分泌的主要粘蛋白。目的探讨自噬在慢性鼻窦炎（CRS）过程中致病性粘蛋白（MUC）5AC产生中的作用。方法检测人中性粒细胞弹性蛋白酶（HNE）及其自噬蛋白的表达。 使用免疫组织化学、酶联免疫吸附测定 (ELISA) 和实时定量分析鼻窦粘膜和人鼻上皮细胞 (HNEC) 中的微管相关蛋白 1 轻链 (LC)3B-II、c-Jun N 末端激酶 (JNK)、c-Jun 和 MUC5AC 聚合酶链反应（qRT-PCR）。使用透射电子显微镜（TEM）研究自噬液泡。用 HNE、巴弗洛霉素 A1 和 SP600125 处理原发性 HNEC。在一些实验中，用小干扰 RNA (siRNA) 转染培养的原代 HNEC，以靶向 Beclin-1 (BECN1；BECN1-siRNA)、自噬相关基因 5 (Atg5；Atg5-siRNA) 和 c-Jun (c-Jun-siRNA)。使用蛋白质印迹、qRT-PCR和ELISA对培养的细胞进行分析。结果在患有或不患有鼻息肉的CRS患者中，HNE、LC3B、JNK、c-Jun和MUC5AC的表达水平上调。 Bafilomycin A1 上调 LC3B-II 表达并抑制 HNE 处理的正常原代 HNEC 中的 MUC 分泌。使用 TEM 在 HNE 处理的原代 HNEC 中观察到自噬体。 BECN1-siRNA、Atg5-siRNA、c-Jun-siRNA 和 SP600125 在正常原代 HNEC 中抑制 HNE 诱导的 MUC5AC 分泌。结论在 HNE 诱导的 CRS 中，自噬通过促进 JNK 和 c-Jun 的磷酸化来增加 MUC5AC 的分泌。
BackgroundAutophagy is a lysosomal degradation pathway that protects the body and is essential for cell survival and differentiation. Mucins (MUCs) are important components of secreted mucus, mucin (MUC)5 AC is the major MUC secreted in the normal airway.ObjectiveInvestigated the role of autophagy in pathogenic mucin (MUC)5 AC production during chronic rhinosinusitis (CRS).MethodsThe expression of human neutrophil elastase (HNE) and the autophagic proteins microtubule-associated protein 1 light chain (LC)3B-II, c-Jun N-terminal kinase (JNK), c-Jun, and MUC5AC were analyzed in the sinonasal mucosa and human nasal epithelial cells (HNECs) using immunohistochemistry, enzyme-linked immunosorbent assay (ELISA), and quantitative real-time polymerase chain reaction (qRT-PCR). Autophagic vacuoles were studied using transmission electron microscopy (TEM). Primary HNECs were treated with HNE, bafilomycin A1, and SP600125. In some experiments, cultured primary HNECs were transfected with small interfering RNAs (siRNAs) to target Beclin-1 (BECN1; BECN1-siRNA), autophagy-related gene 5 (Atg5; Atg5-siRNA), and c-Jun (c-Jun-siRNA). Cultured cells were analyzed using western blotting, qRT-PCR, and ELISA.ResultsIn CRS patients, both with and without nasal polyps, the expression levels of HNE, LC3B, JNK, c-Jun, and MUC5AC were upregulated. Bafilomycin A1 upregulated LC3B-II expression and inhibited MUC secretion in HNE-treated normal primary HNECs. Autophagosomes were observed in HNE-treated primary HNECs using TEM. HNE-induced secretion of MUC5AC was suppressed in normal primary HNECs by BECN1-siRNA, Atg5-siRNA, c-Jun-siRNA, and SP600125.ConclusionsIn HNE-induced CRS, autophagy increases the secretion of MUC5AC by promoting the phosphorylation of JNK and c-Jun.