不可分型流感嗜血杆菌脂肽P4诱导气道上皮细胞分泌黏蛋白的机制  

Nontypeable Haemophilus lipopeptide P4 induces the secretion of MUC5AC in airway epithelial cells

作　　者：王剑[1] 李旎[2] 李晶[2] 周定耕[2] 王彪[2] 

机构地区：[1]南华大学附属第一医院急诊科,衡阳421001 [2]南华大学附属第二医院急诊科

出　　处：《中南医学科学杂志》2017年第4期335-340,共6页Medical Science Journal of Central South China

基　　金：国家自然科学基金(编号:31500156);湖南省卫生厅科研基金(B2014-054)

摘　　要：目的研究不可分型流感嗜血杆菌脂肽P4诱导气道上皮细胞分泌黏蛋白MUC5AC的作用及机制。方法培养人气道上皮细胞NCI-H292,用不同浓度的P4孵育细胞,检测粘蛋白5AC(MUC5AC)的分泌及mRNA表达、ROS的产生及肿瘤坏死因子α转化酶(TACE)的活性;分析Duox1 p47phox和P67phox亚基亚细胞转位和表皮生长因子受体(EGFR)的磷酸化。结果 0、30、50和100 ng/m L P4作用NCI-H292细胞24 h后,可诱导其分泌MUC5AC并表达其mRNA,并促进p47phox和P67phox亚基转位至细胞膜、增高细胞内ROS的含量,同时可上调TACE的酶活性及诱导EGFR磷酸化。NADPH氧化酶抑制剂可抑制ROS产生;而ROS抑制剂处理则可降低TACE的酶活性;沉默TACE表达后可抑制EGFR磷酸化,而EGFR抑制剂处理可降低MUC5AC分泌。结论 P4经Duox1/ROS/TACE/EGFR诱导人NCI-H292细胞分泌MUC5AC。Objective To underly the mechanism of the effect of Nontypeable Haemophilus lipopeptide P4 on the secretion of MUC5AC in airway epithelial cells.MethodsHuman airway epithelial cell line NCI-H292 was incubated with different concentration of P4.The concentrations of secreted MUC5AC and the expression of MUC5AC mRNA,the production of reactive oxygen species （ROS） and the enzymatic activity of tumor necrosis factor-α converting enzyme （TACE） were tested.The cell membrane localization of p47phox and P67phox subunits of Duox1 and the phosphorylation of epidermal growth factor receptor （EGFR） were anylized.Results30,0 and 100 ng/mL of P4 increased the expression and secretion of MUC5AC after 24 h of incubation In addition,P4 could induce the p47phox和P67phox translocation from cytosol to plasma membrane and upregulate the intracellular ROS lever.Moreover,P4 could also promote the enzymatic activity of TACE and phosphorylation of EGFR.Pretreatment of the NADPH oxidase inhibitor significantly abrogated the ROS level,and ROS inhibitor could further decrease the enzymic activity of TACE,while silence of TACE could inhibit P4-induced EGFR phosphorylation.Furthermore,the EGFR inhibitor treatment could decrese the MUC5AC secretion.ConclusionP4 induces the expression and secretion of MUC5AC via Duox1/ROS/TACE/ EGFR signaling pathway in NCI-H292 cells.

关 键 词：不可分型流感嗜血杆菌 脂肽P4 表皮生长因子受体 肿瘤坏死因子α转化酶 粘蛋白5AC 

分 类 号：R56[医药卫生—呼吸系统]