Improved nitric oxide detection using 2,3-diaminonaphthalene and its application to the evaluation of novel nitric oxide synthase inhibitors

Naoki Nakatsubo, Hirotatsu Kojima, Kuniko Sakurai, Kazuya Kikuchi, Hiroshi Nagoshi, Yasunobu Hirata, Takaaki Akaike, Hiroshi Maeda, Yasuteru Urano, Tsunehiko Higuchi, Tetsuo Nagano

研究成果: Article査読

31 被引用数 (Scopus)

抄録

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- tetramethylimidazoline-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 microplate 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.

本文言語English
ページ(範囲)1247-1250
ページ数4
ジャーナルBiological and Pharmaceutical Bulletin
21
12
DOI
出版ステータスPublished - 1998
外部発表はい

ASJC Scopus subject areas

  • 薬理学
  • 薬科学

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