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

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.

本研究的目的是建立高灵敏度的一氧化氮检测系统并应用于临床。本文建立了鲁米诺化学发光法测定一氧化氮的新方法，并应用于大鼠离体灌流肾。使用这个系统，我们发现，内皮素-1释放NO通过刺激ETB受体，这种反应在血管内皮损伤减弱。肾上腺髓质素通过NO-cGMP系统发挥血管舒张作用，其第二信使只有cAMP。加压素通过V1受体增加NO释放。特别是，在动脉血管中，NO可能极大地影响这些血管活性物质在调节血管张力中的作用。我们用臭氧化学发光法分析了肝硬化和肾功能衰竭患者呼出气中的NO排出量，发现NO排出量增加。此外，我们开发了一种对NO特异性敏感的荧光素，并成功地对培养细胞和组织切片中的NO进行了生物成像分析。我们还尝试了一个NO吸入治疗心力衰竭患者。吸入NO可使肺血管扩张，而不改变全身血管系统，导致气体交换增加。此外，NO气体改善了这些患者的运动能力。这些结果表明，吸入NO可能有助于改善心力衰竭患者的生活质量。

项目成果

Ikenouchi H, Hirata Y: "Negative Inotropic Effect of Adrenomedullin in Isolated Adult Rabbit Cardiac Ventricular Myocytes" Circulation. 95. 2318-2324 (1997)

Ikenouchi H、Hirata Y：“肾上腺髓质素对离体成年兔心室肌细胞的负性肌力作用”循环。

Goto Atsuo: "Ouabain-like compound in hypertension associated with ectopic corticotropin syndrome." Hypertension. 28. 421-425 (1996)

Goto Atsuo：“哇巴因样化合物治疗与异位促肾上腺皮质激素综合征相关的高血压。”

Hirata Yasunobu: "Nitric oxide release from kidneys of hyperfensive rats treated with imidapril" Hypertension. (in press). (1996)

Hirata Yasunobu：“用咪达普利治疗的高血压大鼠肾脏释放一氧化氮”高血压。

Hirata Y他: "Receptor subtype for vasopressin-induced release of nitric oxide from rat kidney" Hypertension. 29. 58-64 (1997)

Hirata Y 等人：“加压素诱导大鼠肾脏释放一氧化氮的受体亚型”高血压。 29. 58-64 (1997)

Hirata Yasunobu: "Nitric oxide release from kidneys of hypertensive rats treated with imidapril." Hypertension. 27. 672-678 (1996)

Hirata Yasunobu：“用咪达普利治疗的高血压大鼠肾脏释放一氧化氮。”