文章基本信息

标题：Improved Nitric Oxide Detection Using 2, 3-Diaminonaphthalene and Its Application to the Evaluation of Novel Nitric Oxide Synthase Inhibitors
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作者：Naoki NAKATSUBO ; Hirotatsu KOJIMA ; Kuniko SAKURAI 等
期刊名称：Biological and Pharmaceutical Bulletin
印刷版ISSN：0918-6158
电子版ISSN：1347-5215
出版年度：1998
卷号：21
期号：12
页码：1247-1250
DOI：10.1248/bpb.21.1247
出版社：The Pharmaceutical Society of Japan
摘要：A specific and sensitive detection method for nitric oxide (NO) in living cells and tissue culture systems is required in the search for novel NO synthase (NOS) inhibitors. We have improved a fluorometric determination with 2, 3-diaminonaphthalene (DAN) by the addition of 2-phenyl-4, 4, 5, 5-tetramehylimidazoline-3-oxide-1-oxyl (PTIO) as an oxidant to form NO₂ from NO. This method is 3 times more sensitive than that without PTIO, and is suitable for examining the NOS-inhibitory activity of large numbers of test compounds using a 96-well micropolate reader. The improved method was applied to N-monomethyl-L-arginine (L-NMMA) as a known inhibitor and the derivatives of 2-phenyl-1, 2-benzisoselenazol-3(2H)-one as teat compounds in order to investigate the effect of these compounds on NO production from activated rat aortic smooth muscle cells. The results obtained indicate that this method is suitable for the rapid assay of large numbers of test compounds.
关键词：nitric oxide;inhibitor;nitric oxide synthase;diaminonaphthalene;fluorescence;probe