长期以来,粘蛋白一直被认为是预防粘膜感染的屏障;然而,一些研究报告,粘蛋白的过度表达会引起气道阻塞和炎症。我们调查了是否过度表达的粘蛋白的分泌,粘蛋白5ac(MUC5AC),可以提高对病原体的保护。为了检查粘蛋白高分泌在增强宿主对促进疾病的粘膜阻塞性肺病的防御中的可能作用,产生过表达MUC5AC的小鼠模型。我们以前已经证明,鼠γ疱疹病毒-68(MHV-68)感染可以诱导小鼠肺气肿,后来发展为肺纤维化和肺气肿(CPFE)。我们进一步探索了增加的MUC5AC分泌是否可以提供针对MHV-68诱导的纤维化的益处。我们最初开发了pcDNA3.1-MUC5AC小鼠模型。接下来,实验小鼠随机分为5组:正常对照组,pcDNA3.1对照,pcDNA3.1-MUC5AC,CPFE,和pcDNA3.1-MUC5AC+CPFE。通过苏木精和曙红染色和Masson三色染色对每组进行形态测量分析。通过免疫组织化学染色分析肺组织中的MUC5AC水平,实时聚合酶链反应,和蛋白质印迹分析。通过支气管肺泡灌洗液(BALF)的差异细胞计数和酶联免疫吸附测定BALF中细胞因子和趋化因子的测定来确定气道炎症。仅MUC5AC高分泌不足以驱动杯状细胞化生,从而诱导明显的粘液堵塞和气道炎症。然而,MUC5AC过表达作为体内对抗MHV-68病毒感染的保护性屏障。与CPFE组相比,pcDNA3.1-MUC5ACCPFE组的MHV-68感染性降低。同时,MHV-68病毒的减少减弱了趋化因子(C-C基序)配体2(CCL2)的表达,趋化因子(C-X-C基序)配体5(CXCL5),白细胞介素-13(IL-13),和转化生长因子-β1(TGF-β1),pcDNA3.1-MUC5AC+CPFE组气道炎症和纤维化减弱。MUC5AC的过表达似乎在随后发展为CPFE的肺气肿小鼠中表现出对MHV-68感染的保护作用,并通过降低CCL2,CXCL5,IL-13和TGF-β1的表达进一步降低MHV-68诱导的气道炎症和纤维化。
Mucins have long been regarded to play a role as a barrier to prevent mucosal infections; however, some studies report that overexpression of mucins induces obstruction and inflammation of airways. We investigated whether the secretion of overexpressed mucin, mucin5ac (MUC5AC), could improve protection against pathogens. To examine the possible roles of mucin hypersecretion in augmenting host defense against disease-promoting muco-obstructive lung disease, a mouse model that overexpressed MUC5AC was generated. We had previously proved that murine gammaherpesvirus-68 (MHV-68) infection could induce emphysema in mice, which later developed into combined pulmonary fibrosis and emphysema (CPFE). We further explored whether increased MUC5AC secretion could provide benefits against MHV-68 induced fibrosis. We initially developed a pcDNA3.1-MUC5AC mouse model. Next, the experimental mice were randomly divided into five groups: normal control, pcDNA3.1 control, pcDNA3.1-MUC5AC, CPFE, and pcDNA3.1- MUC5AC + CPFE. Morphometric analysis of each group was performed by hematoxylin and eosin staining and Masson trichrome staining. MUC5AC levels in lung tissues were analyzed by immunohistochemical staining, real-time polymerase chain reaction, and Western blot analysis. The airway inflammation was determined by differential cell counts of bronchoalveolar lavage fluid (BALF) and measurement of cytokines and chemokines in BALF by enzyme-linked immunosorbent assay. MUC5AC hypersecretion alone was not sufficient to drive goblet cell metaplasia to induce obvious mucus plugging and airway inflammation. However, MUC5AC overexpression served as a protective barrier against MHV-68 virus infection in vivo. Infectivity of MHV-68 was decreased in the pcDNA3.1-MUC5AC + CPFE group compared with that in CPFE group. Meanwhile, a reduction of MHV-68 virus attenuated the expressions of chemokine (C-C motif) ligand 2 (CCL2), chemokine (C-X-C motif) ligand 5 (CXCL5), interleukin-13 (IL-13), and transforming growth factor-β1 (TGF-β1), and weakened airway inflammation and fibrosis in the pcDNA3.1-MUC5AC + CPFE group. Overexpression of MUC5AC appears to exhibit a protective role against MHV-68 infection in mice with emphysema that subsequently developed into CPFE and to further decrease airway inflammation and fibrosis induced by MHV-68 by decreasing the expressions of CCL2, CXCL5, IL-13, and TGF-β1.