使用自由基清除剂羧基 PTIO 和格里斯试剂，改进对活化巨噬细胞培养物中一氧化氮自由基（NO.）产生的检测。

Improved detection of nitric oxide radical (NO.) production in an activated macrophage culture with a radical scavenger, carboxy PTIO and Griess reagent.

作者信息

Amano F, Noda T

机构信息

Department of Biochemistry and Cell Biology, National Institute of Health, Tokyo, Japan.

出版信息

FEBS Lett. 1995 Jul 24;368(3):425-8. doi: 10.1016/0014-5793(95)00700-j.

DOI:10.1016/0014-5793(95)00700-j

PMID:7543422

Abstract

An improved method for the detection of nitric oxide radicals (NO. in cultures of activated macrophages was developed, involving a nitric oxide radical scavenger, 2-(4-carboxyphenyl)-4,4,5,5- tetramethylimidazoline-3-oxide-1-oxyl (carboxy PTIO) and Griess reagent. A murine macrophage-like cell line, J774.1, was activated with interferon-gamma (IFN-gamma) and bacterial lipopolysaccharide (LPS), which induced the production and secretion of NO2- into the culture supernatant. Addition of carboxy PTIO to the activated macrophages increased the amount of NO2- to 4- to 5-fold without cell damages, probably because carboxy PTIO rapidly reacted with NO. to form NO2-, which was finally assayed by the Griess reaction.

摘要

开发了一种改进的方法用于检测活化巨噬细胞培养物中的一氧化氮自由基（NO·），该方法涉及一氧化氮自由基清除剂2-(4-羧基苯基)-4,4,5,5-四甲基咪唑啉-3-氧化物-1-氧基（羧基PTIO）和格里斯试剂。用γ干扰素（IFN-γ）和细菌脂多糖（LPS）激活小鼠巨噬细胞样细胞系J774.1，诱导其产生并分泌NO2-到培养上清液中。向活化的巨噬细胞中添加羧基PTIO可使NO2-的量增加4至5倍且无细胞损伤，这可能是因为羧基PTIO与NO·迅速反应形成NO2-，最终通过格里斯反应进行测定。