Airway epithelical Cl channel regulation by iNOS gene

iNOS 基因对气道上皮 Cl 通道的调节

基本信息

批准号：
10670563
负责人：
TAMAOKI Jun
金额：
$ 1.92万
依托单位：
Tokyo Women's Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10670563/
关键词：
Airway epithelium Nitric oxide Ion channel Patch clamp Gene expression Airway secretion Airway inflammation Asthma 気道分必

项目摘要

There is increasing evidence that nitric oxide (NO) is synthesized and released in the airways, thereby playing a role in a variety of airway functions. In the present experiment, to determine whether epithelium-derived NO is involved in the regulation of Cl secretion and, hence, water secretion into the airway lumen, we studied isolated human bronchial epithelial cells in vitro. Patch clamp studies of isolated airway epithelium showed that intracellular adition of high calcium solution plus bradykinin, or the NO donors S-nitroacetyl-N-penicillamine and nitroprusside caused an increase in Cl-selective current and open probability of luminal Cl channel. This effect was accompanied by the corresponding increase in NO-selective electrical current, as measured by a polarography using an NO-selective electrode, and induction of the presence of constitutive NO synthase (cNOS)-like immunoreactivities. On the other hand, stimulation of the cells with endotoxin from E. coli, interleukin-1β, tumor necrosis factor-α, or interferon-γ each increased Cl conductance and single Cl channel current. This effect was likewise accompanied by the increase in NO concentration in the medium, but, based on Nothern analysis and immunocytochemistry, expressions of iNOS mRNA and iNOS protein were upregulated by these stimuli. Futhermore,these effects were dose-dependently inhibited by 14-membered macrolides and glucocorticosteroids. Therefore, macrolides and steroids may be therapeutic potential for the treatment of airway hypersecretion in the inflamed airways.

越来越多的证据表明，一氧化氮（NO）在气道中合成并释放，从而在各种气道功能中发挥作用。在演讲实验中，为了确定上皮衍生的NO是否参与了CL分泌的调节，因此，我们在体外研究了隔离的人支气管上皮细胞。对隔离气道上皮的贴片夹研究表明，高钙溶液加上心动激肽的细胞内版本，或无供体S-硝基乙酰基-N-苯胺和硝化剂导致Cl-selesextive电流的增加和腔内CL通道的开放概率。这种效果伴随着通过使用无选择电极的警察摄影测量的无选择的电流的相应增加，以及诱导本构型NO合酶（CNOS）类似于类似于的免疫反应性的存在。另一方面，用大肠杆菌，白细胞介素1β，肿瘤坏死因子-α或干扰素γ的细胞刺激细胞，每个都会增加Cl电导量和单个CL通道电流。这种效果同样是通过培养基中无浓度的增加来实现的，但是，基于Nothern分析和免疫细胞化学，这些刺激更新了Inos mRNA和Inos蛋白的表达。 festhermore，这些作用被14元的大环内酯类和糖皮质激素依赖性抑制。因此，大环内酯类和类固醇可能是在发炎的气道中治疗气道突出分泌的热潜力。