Kikuchi K, Nagano T, Hayakawa H, Hirata Y, Hirobe M
Faculty of Pharmaceutical Sciences, University of Tokyo, Japan.
Anal Chem. 1993 Jul 1;65(13):1794-9. doi: 10.1021/ac00061a025.
Nitric oxide, thought to be the endothelium-derived relaxing factor (EDRF), is involved in intra- and intercellular signalling in various tissues. A system for the continuous detection of NO in the picomolar range from a perfused organ is described. The detection is based upon the chemiluminescence reaction between NO and the luminol (5-amino-2,3-dihydro-1,4-phthalazinedione)-H2O2 system. The chemiluminescence is due to the formation of peroxynitrite from NO and H2O2. The luminol-H2O2 system is specifically reactive to NO, so that other nitrogen-containing compounds, (organic nitrite, organic nitrate, and thio-nitroso compounds) or endothelium-derived compounds do not interfere. The limit of determination was approximately 100 fM. This system has been used to measure continuous NO release from isolated perfused rat kidney and the simultaneous changes of perfusion pressure. In Wistar rats basal NO release was 85 +/- 11 fmol/min-1 (g of kidney weight)-1 (39 pM in the perfusate), and acetylcholine increased NO release dose dependently with a concomitant pressure reduction. The changes in NO release were always associated with mirror image changes in the perfusion pressure. Simple pressure reduction did not interfere with the chemiluminescence. Precise titration data as well as results of some preliminary experiments using this method are presented.
一氧化氮被认为是内皮衍生舒张因子（EDRF），参与多种组织的细胞内和细胞间信号传导。本文描述了一种用于连续检测灌注器官中皮摩尔范围内一氧化氮的系统。该检测基于一氧化氮与鲁米诺（5-氨基-2,3-二氢-1,4-酞嗪二酮）-过氧化氢系统之间的化学发光反应。化学发光是由于一氧化氮和过氧化氢形成过氧亚硝酸盐所致。鲁米诺-过氧化氢系统对一氧化氮具有特异性反应，因此其他含氮化合物（有机亚硝酸盐、有机硝酸盐和硫代亚硝基化合物）或内皮衍生化合物不会产生干扰。测定限约为100飞摩尔。该系统已用于测量离体灌注大鼠肾脏中一氧化氮的持续释放以及灌注压力的同步变化。在Wistar大鼠中，基础一氧化氮释放量为85±11飞摩尔/分钟-1（每克肾脏重量）-1（灌注液中为39皮摩尔），乙酰胆碱可剂量依赖性地增加一氧化氮释放，并伴有压力降低。一氧化氮释放的变化总是与灌注压力的镜像变化相关。单纯的压力降低不会干扰化学发光。本文给出了精确的滴定数据以及使用该方法的一些初步实验结果。
