Cigarette smoke extract activates human bronchial epithelial cells affecting non-neuronal cholinergic system signalling in vitro
Cigarette smoke extract activates human bronchial epithelial cells affecting non-neuronal cholinergic system signalling in vitro
复制标题
DOI：
10.1016/j.lfs.2011.04.025
复制
发表时间：
2011-07-04
期刊：
影响因子：
6.1
通讯作者：
Gjomarkaj, Mark
中科院分区：
文献类型：
--
作者：
Profita, Mirella;Bonanno, Anna;Gjomarkaj, Mark
目的：乙酰胆碱（Acetylcholine，ACh）是由胆碱乙酰转移酶（Choline Acetyl-Transferase，ChAT）合成的，ChAT通过激活毒蕈碱受体（Muscarinic Receptor，MR）发挥其在气道上皮细胞中的生理作用。我们评估ACh刺激对人支气管上皮细胞（16-HBE）的影响，并测试香烟烟雾提取物（CSE）是否可以改变对ACh的基础细胞反应，从而影响非神经元胆碱能系统signaling.Main方法：ACh刺激的16-HBE进行ACh结合测试。白三烯B-4（LTB 4）释放和ERK 1/2和NF κ B通路激活。此外，我们研究了所有上述参数以及ChAT和MR蛋白和mRNA的表达和内源性ACh的生产在CSE处理16-HBE.Key结果：我们表明，ACh诱导16-HBE，in浓度依赖性的方式，LTB 4的释放通过激活ERK 1/2和NF κ B途径。添加噻托溴铵（思力华（R））、没食子胺、替仑西平和4-DAMP（毒蕈碱受体拮抗剂）以及PD 098059（MAPKK抑制剂）和BAY 117082（1 kB α磷酸化抑制剂）可下调乙酰胆碱诱导的效应。此外，CSE处理16-HBE增加了ACh的结合，并将LTB 4的释放从ACh浓度1 μ M转移到10 nM。最后，我们观察到用CSE处理16-HBE增加了ChAT、M-2和M-3的表达以及16-HBE中内源性ACh的产生。噻托溴铵可调节CSE处理的16-HBE中LTB 4的释放和ACh的产生。意义：CSE通过影响ChAT和MRS的表达，增强人支气管上皮细胞的促炎活性，促进细胞对低浓度ACh的反应。噻托溴铵可能阻止ACh与CSE共同产生的促炎事件。(C)2011 Elsevier Inc. All rights reserved.
Aims: Acetylcholine (ACh) is synthesized by Choline Acetyl-Transferase (ChAT) that exerts its physiological effects in airway epithelial cells via muscarinic receptor (MR) activation. We evaluate the effect of ACh stimulation on human bronchial epithelial cells (16-HBE) and test whether cigarette smoke extract (CSE) can modify the basal cellular response to ACh affecting the non-neuronal cholinergic system signalling.Main methods: ACh stimulated 16-HBE were tested for ACh-binding. Leukotriene B-4 (LTB4) release and ERK1/2 and NFkB pathway activation. Additionally, we investigated all the aforementioned parameters as well as ChAT and MR proteins and mRNA expression and endogenous ACh production in CSE-treated 16-HBE.Key findings: We showed that ACh induced in 16-HBE, in a concentration-dependent manner, LTB4 release via the activation of ERK1/2 and NFkB pathways. The addition of Tiotropium (Spiriva (R)), Gallamine, Telenzepine and 4-DAMP (muscarinic receptor antagonists), as well as of PD 098059 (MAPKK inhibitor) and BAY117082 (inhibitor of 1kB alpha phosphorilation), down-regulated the ACh-induced effects. Additionally, CSE treatment of 16-HBE increased the binding of ACh, and shifted the LTB4 release from the concentration ACh 1 mu M to 10 nM. Finally, we observed that the treatment of 16-HBE with CSE increased the expression of ChAT, M-2 and M-3 and of endogenous ACh production in 16-HBE. Tiotropium regulated the LTB4 release and ACh production in CSE treated 16-HBE.Significance: CSE increases the pro-inflammatory activity of human bronchial epithelial cells, and promotes the cellular response to lower concentrations of ACh, by affecting the expression of ChAT and MRs. Tiotropium might prevent pro-inflammatory events generated by ACh together with CSE. (C) 2011 Elsevier Inc. All rights reserved.