Cold temperature induces mucin hypersecretion from normal human bronchial epithelial cells in vitro through a transient receptor potential melastatin 8 (TRPM8)-mediated mechanism
Cold temperature induces mucin hypersecretion from normal human bronchial epithelial cells in vitro through a transient receptor potential melastatin 8 (TRPM8)-mediated mechanism
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低温通过瞬态受体电位melastatin 8 (TRPM8)介导的机制在体外诱导正常人支气管上皮细胞粘蛋白过度分泌
DOI：
10.1016/j.jaci.2011.04.032
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发表时间：
2011-09-01
影响因子：
14.2
通讯作者：
Zhou, Xiang Dong
中科院分区：
文献类型：
--
作者：
Li, MinChao;Li, Qi;Zhou, Xiang Dong
背景资料：冷空气刺激是加重慢性炎症性气道疾病（如慢性阻塞性肺疾病（COPD）和哮喘）的主要环境因素。在分子水平上，冷通过瞬时受体电位melastatin 8（TRPM 8）检测。迄今为止，TRPM 8表达尚未在COPD患者的气道上皮中表征。TRPM 8通道在冷刺激诱导的一系列气道反应中的作用以及TRPM 8在调节冷诱导反应中的分子和生化途径在很大程度上是未知的。目的：我们试图探索TRPM 8在冷空气引起的粘液高分泌中的作用以及参与该过程的潜在信号通路。采用免疫组化、RT-PCR和Western blotting方法检测支气管上皮中TRPM 8的表达。用钙离子成像和磷脂酶C（PLC）δ 1-pleckstrin同源物-绿色荧光蛋白时空动力学方法分别表征TRPM 8受体功能和磷脂酰肌醇4，5-二磷酸（PIP 2）水解。实时荧光定量PCR法检测MUC 5AC mRNA的表达，ELISA法检测MUC 5AC粘蛋白的表达。在豆蔻酰化富含丙氨酸的C激酶底物（MARCKS）的磷酸化位点域的四个丝氨酸残基突变，以确定TRPM 8介导的气道粘液hypersecretion.Results的MARCKS的功能：TRPM 8蛋白和mRNA的表达显着增加，COPD患者的表达相比，在健康受试者。低温导致细胞内Ca（2+）水平显著升高，并促进PLC delta 1-pleckstrin同源物-绿色荧光蛋白的转位。冷刺激可非持续性地增加MUC 5AC mRNA和细胞内及分泌的MUC 5AC蛋白的表达。结论：TRPM 8受体通过Ca（2+）-PLC-PIP 2-MARCKS信号通路参与冷诱导的粘液高分泌。（J Allergy Clin Immunol 2011;128：626-34.）
Background: Cold air stimulus is a major environmental factor that exacerbates chronic inflammatory airway diseases, such as chronic obstructive pulmonary disease (COPD) and asthma. At the molecular level, cold is detected by transient receptor potential melastatin 8 (TRPM8). To date, TRPM8 expression has not been characterized in the airway epithelium of patients with COPD. The role of TRPM8 channels in a series of airway responses induced by cold stimuli and the molecular and biochemical pathways of TRPM8 in regulating cold-induced responses are largely unknown.Objective: We sought to explore the role of TRPM8 in cold air-provoked mucus hypersecretion and the potential signaling pathway involved in this process.Methods: The expression of TRPM8 in the bronchial epithelium was examined by means of immunohistochemistry, RT-PCR, and Western blotting. TRPM8 receptor function and hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP2) were characterized by means of Ca(2+) imaging and spatiotemporal dynamics of phospholipase C (PLC) delta 1-pleckstrin homology-green fluorescent protein, respectively. The expression of MUC5AC mRNA and MUC5AC mucin protein was measured by using real-time PCR and ELISA, respectively. Four serine residues in the myristoylated alanine-rich C kinase substrate (MARCKS)-phosphorylation site domain were mutated to identify the function of MARCKS in TRPM8-mediated airway mucus hypersecretion.Results: TRPM8 protein and mRNA expression were significantly increased in patients with COPD compared with expression seen in healthy subjects. Cold produced robust increases in intracellular Ca(2+) levels and promoted translocation of PLC delta 1-pleckstrin homology-green fluorescent protein. Cold increased expression of MUC5AC mRNA and intracellular and secreted MUC5AC protein in a nonsustained way. Phosphorylation site domain-mutant MARCKS cDNA hindered MUC5AC secretion induced by cold.Conclusions: These results indicate that the TRPM8 receptor is involved in cold-induced mucus hypersecretion through the Ca(2+)-PLC-PIP2-MARCKS signaling pathway. (J Allergy Clin Immunol 2011;128:626-34.)