An assay method for nitric oxide-related compounds in whole blood.
An assay method for nitric oxide-related compounds in whole blood.
复制标题
全血中一氧化氮相关化合物的测定方法。
作者：
M. Sonoda;J. Kobayashi;M. Takezawa;T. Miyazaki;T. Nakajima;H. Shimomura;K. Koike;A. Satomi;H. Ogino;R. Omoto;T. Komoda
最近的证据表明，体内产生的一氧化氮 (NO) 在循环中会转化为亚硝酸盐/硝酸盐、亚硝酰血红蛋白、亚硝酰金属复合物和 S-亚硝基化合物的形式。据报道，亚硝基硫醇是相对稳定的代谢物，在血浆中具有微摩尔水平。因此，我们假设血液中所有 NO 相关化合物的测定都具有诊断意义。这里描述的测定方法包括全血中所有 NO 相关化合物的热解以及酶促还原后通过荧光测定或化学发光检测所得硝酸盐。 S-亚硝基白蛋白和亚硝酰血红蛋白很容易热解为硝酸盐，相对稳定的S-亚硝基谷胱甘肽在高分子量（30 kDa以上）血液成分存在下也会降解为硝酸盐。测定了健康人以及对照或脂多糖刺激的大鼠血液中 NO 相关化合物的浓度。我们发现膜结合的NO在病理生理状态下表现出增强的水平也可以被检测到。结合电子自旋共振光谱，我们的数据表明，红细胞内扩散和代谢的NO分数以及血浆中的其他NO代谢物，如亚硝酸盐/硝酸盐和S-亚硝基化合物，两者都可以反映体内NO的产生，可以通过该方法进行回收和定量检测。
Recent evidence suggests that nitric oxide (NO) generated in vivo will be converted into the forms of nitrite/nitrate, nitrosyl hemoproteins, nitrosyl metal complexes, and S-nitroso-compounds in the circulation. Nitrosothiols have also been reported to be relatively stable metabolites with micromolar levels in plasma. We hypothesized, therefore, that the determinations of all the NO-related compounds in blood would be of diagnostic significance. The assay method described here consists of the thermolysis of all the NO-related compounds in whole blood and the detection of resulting nitrate by fluorometry or chemiluminescence after an enzymatic reduction. S-Nitroso-albumin and nitrosyl hemoglobin can be easily thermolysed to nitrate, and relatively stable S-nitroso-glutathione is also degraded to nitrate in the presence of blood constituents with high molecular mass (above 30 kDa). Concentrations of NO-related compounds in blood from healthy human as well as control or lipopolysaccharide-stimulated rats were determined. We found that membrane-bound NO which showed the augmented levels under the pathophysiological states could also be detected. Together with electron spin resonance spectra, our data indicate that the fraction of NO diffused and metabolized within red cells and the other NO-metabolites in plasma such as nitrite/nitrate and S-nitroso-compounds, both of which can reflect NO-production in vivo, would be recovered and detected quantitatively by this method.
登录
查看更多内容
影响因子：
3.9
作者：
ARNELLE, DR;STAMLER, JS
通讯作者：
STAMLER, JS
动脉粥样硬化兔胆碱能小动脉血管舒张缺陷的视频显微镜演示。
DOI：
10.1172/jci113516
发表时间：
1988
期刊：
The Journal of clinical investigation
影响因子：
--
作者：
Yamamoto,H;Bossaller,C;CartwrightJr,J;Henry,PD
通讯作者：
Henry,PD
DOI：
10.1016/s0021-9258(18)54642-1
发表时间：
1991-11
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
J. A. Corbett;Jack R. Lancaster;M. Sweetland;M. L. McDaniel
通讯作者：
J. A. Corbett;Jack R. Lancaster;M. Sweetland;M. L. McDaniel
人肝细胞中诱导型一氧化氮合酶的分子克隆和表达
DOI：
10.1073/pnas.90.8.3491
发表时间：
1993-04-15
影响因子：
11.1
作者：
GELLER, DA;LOWENSTEIN, CJ;BILLIAR, TR
通讯作者：
BILLIAR, TR
DOI：
10.1073/pnas.89.16.7674
发表时间：
1992-08-15
影响因子：
11.1
作者：
STAMLER, JS;JARAKI, O;LOSCALZO, J
通讯作者：
LOSCALZO, J