Development of sensitve assay of NO and its clinical application

NO灵敏检测方法的研制及其临床应用

• 批准号: 07557055
• 负责人: HIRATA Yasunobu
• 金额: $ 6.98万
• 依托单位: University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07557055/
• 关键词: nitric oxide; chemiluminescence; inhalation; exhaled air; arteriosclerosis; kidney; 化学発光; NO合成酵素; nitric oxide; chemiluminescence; inhalation; exhaled air; arteriosclerosis; kidney; 化学発光; NO合成酵素

The aims of the present study were development of highly sensitive NO assay system and its clinical application. We newly developed luminol chemiluminescence assay for NO and applied it to the rat isolated perfused kidney. Using this system, we found that endothelin-1 released NO through stimulation of ETB receptors and that this response was attenuated in the vessels with endothelial damage. Adrenomedullin, of which second messenger had been believed only cAMP,exhibited vasodilatory action by NO-cGMP system. Vasopressin increased NO release through V1 receptors. In particular, in arteriosclerotic vessels, NO may greatly influence a role of these vasoactive substances in regulation of vascular tone. We analyzed NO output in the exhaled air of patients with liver cirrhosis and renal failure by ozone chemiluminescence, and found the increase in NO excretion. Furthermore, we developed a fluorescein specifically sensitive to NO,and succeeded in bioimaging analysis of NO in the cultured cells and tissue slices. We also tried an NO inhalation therapy in patients with heart failure. Inhaled NO dilated pulmonary vessels without changes in systemic vasculature, resulting in an increase in gas exchange. Moreover, NO gas improved exercise capacity in these patients. These results suggest that NO inhalation may be useful for improvement of quality of life in heart failure.

本研究的目的是开发高度敏感的NO分析系统及其临床应用。我们对NO的新开发了Luminol化学发光测定法，并将其应用于大鼠孤立的灌注肾脏。使用该系统，我们发现内皮素-1通过刺激ETB受体释放NO，并且这种反应在血管中受到内皮损伤的衰减。肾上腺素甲素（肾上腺素）仅被认为只有营地，它表现出No-CGMP系统的血管舒张作用。加压素通过V1受体增加了无释放。特别是，在动脉硬化血管中，没有可能会极大地影响这些血管活性物质在调节血管张力中的作用。我们通过臭氧化学发光的肝硬化和肾衰竭患者的呼出空气中没有分析输出，发现无排泄的增加。此外，我们开发了一种对NO特别敏感的荧光素，并在培养的细胞和组织切片中成功地分析了NO的生物成像分析。我们还尝试了心力衰竭患者的无吸入疗法。吸入未扩张的肺部血管，没有系统性脉管系统的变化，从而导致气体交换增加。此外，这些患者没有气体提高运动能力。这些结果表明，没有吸入可能对改善心力衰竭生活质量可能有用。