Improved nitric oxide detection using 2,3-diaminonaphthalene and its application to the evaluation of novel nitric oxide synthase inhibitors
| Autor: | Hiroshi Nagoshi, Naoki Nakatsubo, Hirotatsu Kojima, Yasunobu Hirata, Takaaki Akaike, Tetsuo Nagano, Yasuteru Urano, Kuniko Sakurai, Kazuya Kikuchi, Tsunehiko Higuchi, Hiroshi Maeda |
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| Rok vydání: | 1999 |
| Předmět: |
Azoles
Male Fluorescence spectrometry Pharmaceutical Science In Vitro Techniques Isoindoles Nitric Oxide Muscle Smooth Vascular Nitric oxide Tissue culture chemistry.chemical_compound 2-Naphthylamine Organoselenium Compounds Animals Enzyme Inhibitors Rats Wistar Aorta Pharmacology omega-N-Methylarginine biology Biological activity General Medicine In vitro Rats Nitric oxide synthase chemistry Biochemistry Enzyme inhibitor biology.protein Omega-N-Methylarginine Nitric Oxide Synthase |
| Zdroj: | Biologicalpharmaceutical bulletin. 21(12) |
| ISSN: | 0918-6158 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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