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Nitric oxide Production and regulation in airway epithelium measured by microsensor

微传感器测量气道上皮中一氧化氮的产生和调节

基本信息

批准号：
07670680
负责人：
KONDO Mitsuko
金额：
$ 0.38万
依托单位：
Tokyo Women's Medical College
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07670680/
关键词：
Nitric Oxide airway epithelial cells nitric oxide synthase calcium cyclic AMP cyclic GMP Chloride ion ion transport

项目摘要

Nitric oxide (NO) is produced from various types of cells including airway epithelial cells and plays an important role in physiologic and inflammatory processes in the airway. In this study, NO production from cultured epithelial cells was measured at real time by NO selective microsensor, polarographically. This study was aimed to determine the role of NO in the airway epithelium.1.NO production from airway epitheliumAirway epithelial cells from cow trachea produced NO spontaneously. Isoproterenol (ISO) and dibutylyl cyclic AMP stimulated NO production. This response was inhibitted by L-NG-nitroarginine methylester (L-NAME). In contrast, ATP and bradykinin (BK) had little effect on NO production. These data suggest that NO production from airway epithelium is mediated via cyclic AMP.2.The effect of NO on Ca^<2+> dynamicsL-NAME did not change baseline level of intracellular calcium ([Ca^<2+>]i). However, pretreatment of the cells with L-NAME,but not D-NAME,inhibited ATP-and BK-induced increase in [Ca^<2+>]i. This inhibitory effect was reversed by L-arginine.Furthermore, pretreatment of the cells with nitroprusside and dibutyryl cyclic GMP potentiated ATP-and BK-induced increase in [Ca^<2+>]i. These results suggest that endogenous and exogenous NO affect Ca^<2+> dynamics in the airway epithelium.3.The effect of NO on Cl secretionThe cell sheets ware mounted in Ussing chamber in the presence of amiloride to assess active Cl secretion. Short circuit current (Isc) was continuously monitored. L-NAME alone induced little decrease in Isc. Pretreatment of the cells with L-NAME,but not D-NAME,strongly inhibited ATP-, BK-and ISO-induced increases in Isc. This L-NAME-induced suppression was dose-dependent and reversed by the simultaneous addition of L-arginine. These data suggest that endogenous NO is a key molecule for agonist-induced Cl secretion.

一氧化氮（NO）由包括气道上皮细胞在内的多种细胞产生，在气道的生理和炎症过程中起重要作用。在这项研究中，通过一氧化氮选择性微传感器，用极谱法实时测量培养上皮细胞的一氧化氮产量。本研究旨在探讨一氧化氮在气道上皮中的作用。牛气管上皮细胞自发产生NO。异丙肾上腺素（ISO）和二丁基环AMP刺激NO的产生。这种反应被l - ng -硝基精氨酸甲基lester （L-NAME）抑制。相比之下，ATP和缓激素（BK）对NO的产生影响不大。这些数据表明气道上皮一氧化氮的产生是通过环AMP.2介导的。NO对Ca^<2+>动态的影响没有改变细胞内钙的基线水平（[Ca^<2+>]i）。然而，用L-NAME而不是D-NAME预处理细胞，可以抑制atp和bk诱导的[Ca^<2+>]i的增加。这种抑制作用被l -精氨酸逆转。此外，硝普苷和二丁基环GMP预处理细胞可增强atp和bk诱导的[Ca^<2+>]i升高。这些结果表明，内源性和外源性NO影响气道上皮内Ca^<2+>的动态。一氧化氮对氯离子分泌的影响将细胞片置于氨酰氯存在的Ussing室中，以评估活跃的氯离子分泌。连续监测短路电流（Isc）。单独使用L-NAME对Isc的影响较小。用L-NAME而非D-NAME预处理细胞，能强烈抑制ATP-、bk和iso诱导的Isc升高。这种l - name诱导的抑制是剂量依赖性的，同时加入l -精氨酸可以逆转。这些数据表明内源性NO是激动剂诱导的Cl分泌的关键分子。