Nitric oxide synthetic pathway in red blood cells is impaired in coronary artery disease.
BACKGROUND
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.


METHODS
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-[(15)N2]arginine conversion to L-[(15)N]citrulline respectively.


RESULTS
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.


CONCLUSIONS
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.
Centro Cardiologico Monzino, Istituto di Ricovero e Cura a Carattere Scientifico (I.R.C.C.S.), Milan, Italy.
Sonia
S
Eligini
Benedetta
B
Porro
Alessandro
A
Lualdi
Isabella
I
Squellerio
Fabrizio
F
Veglia
Elisa
E
Chiorino
Mauro
M
Crisci
Anna
A
Garlaschè
Marta
M
Giovannardi
Josè-Pablo
JP
Werba
Elena
E
Tremoli
Viviana
V
Cavalca
D000368:Aged; D001120:Arginine / Q000031:analogs & derivatives / Q000097:blood / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D003324:Coronary Artery Disease / Q000097:blood* / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism*; D005260:Female; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D008297:Male; D008875:Middle Aged; D009569:Nitric Oxide / Q000097:blood* / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000097:blood / Q000378:metabolism; D018384:Oxidative Stress / Q000502:physiology; D053719:Tandem Mass Spectrometry
D009569:Nitric Oxide; C024917:symmetric dimethylarginine; D001120:Arginine; D019001:Nitric Oxide Synthase; D005978:Glutathione