Nitric oxide synthetic pathway in red blood cells is impaired in coronary artery disease.
Nitric oxide synthetic pathway in red blood cells is impaired in coronary artery disease.
复制标题
DOI：
10.1371/journal.pone.0066945
复制
发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Cavalca V
中科院分区：
文献类型：
--
作者：
Eligini S;Porro B;Lualdi A;Squellerio I;Veglia F;Chiorino E;Crisci M;Garlaschè A;Giovannardi M;Werba JP;Tremoli E;Cavalca V
所有参与一氧化氮（NO）代谢的酶促因子/辅助因子都是最近在红细胞中发现的。增加的氧化应激损害NO的生物利用度，并已在冠状动脉疾病（CAD）患者的血浆中被描述。该研究的目的是强调与健康对照相比，冠心病患者红细胞和血浆中NO代谢谱的潜在功能障碍。采用液相色谱串联质谱法和高效液相色谱法测定l -精氨酸/NO途径。采用液相色谱串联质谱法测定谷胱甘肽氧化形态和还原形态的比值，作为氧化应激的指标。通过免疫荧光染色和L-[15N2]精氨酸转化为L-[15N]瓜氨酸的离体实验，分别评价NO合成酶的表达和活性。在冠心病患者的红细胞和血浆中，不对称和对称二甲基精氨酸的含量均比对照组增加。有趣的是，NO合成酶的表达和活性在CAD红细胞中降低。相反，氧化/还原性谷胱甘肽比率在冠心病中升高，并与精氨酸酶活性有关。我们的研究首次分析了l -精氨酸/NO在红细胞和血浆中的整个代谢途径，强调了CAD患者红细胞中NO途径的损伤，与NO合成酶表达/活性降低和氧化应激增加有关。
All the enzymatic factors/cofactors involved in nitric oxide (NO) metabolism have been recently found in red blood cells. Increased oxidative stress impairs NO bioavailability and has been described in plasma of coronary artery disease (CAD) patients. The aim of the study was to highlight a potential dysfunction of the metabolic profile of NO in red blood cells and in plasma from CAD patients compared with healthy controls. We determined L-arginine/NO pathway by liquid-chromatography tandem mass spectrometry and high performance liquid chromatography methods. The ratio of oxidized and reduced forms of glutathione, as index of oxidative stress, was measured by liquid-chromatography tandem mass spectrometry method. NO synthase expression and activity were evaluated by immunofluorescence staining and ex-vivo experiments of L-[15N2]arginine conversion to L-[15N]citrulline respectively. Increased amounts of asymmetric and symmetric dimethylarginines were found both in red blood cells and in plasma of CAD patients in respect to controls. Interestingly NO synthase expression and activity were reduced in CAD red blood cells. In contrast, oxidized/reduced glutathione ratio was increased in CAD and was associated to arginase activity. Our study analyzed for the first time the whole metabolic pathway of L-arginine/NO, both in red blood cells and in plasma, highlighting an impairment of NO pathway in erythrocytes from CAD patients, associated with decreased NO synthase expression/activity and increased oxidative stress.
登录
查看更多内容
影响因子：
3.7
作者：
Cavalca V;Veglia F;Squellerio I;De Metrio M;Rubino M;Porro B;Moltrasio M;Tremoli E;Marenzi G
通讯作者：
Marenzi G
影响因子：
10.8
作者：
Jung, Christian;Gonon, Adrian T.;Pernow, John
通讯作者：
Pernow, John
影响因子：
20.3
作者：
Kleinbongard, P;Schulz, R;Kelm, M
通讯作者：
Kelm, M
DOI：
10.1007/978-1-61779-867-2_21
发表时间：
2012-01-01
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
Harris, Craig;Hansen, Jason M
通讯作者：
Hansen, Jason M
DOI：
10.1152/ajpheart.00066.2006
发表时间：
2006-10-01
影响因子：
4.8
作者：
Billecke, Scott S.;Kitzmiller, Laura A.;D'Alecy, Louis G.
通讯作者：
D'Alecy, Louis G.