Expression of an endothelial-type nitric oxide synthase isoform in human neutrophils: modification by tumor necrosis factor-alpha and during acute myocardial infarction.
OBJECTIVE
The purpose of this study was to determine whether human neutrophils express an endothelial-type nitric oxide synthase (eNOS), and to study the effect of tumor necrosis factor-alpha (TNF-alpha) on its expression.


BACKGROUND
Several studies have demonstrated the presence of a constitutively expressed nitric oxide svnthase (NOS) in neutrophils. Cardiovascular disease is characterized by increased levels of plasma TNF-alpha, a cytokine that has demonstrated eNOS messenger ribonucleic acid (mRNA) destabilization in cultured endothelial cells.


METHODS
Neutrophils were obtained from healthy volunteers and from patients with acute myocardial infarction (AMI).


RESULTS
Human neutrophils express eNOS mRNA and eNOS protein. Stimulation of neutrophils with TNF-alpha decreased eNOS protein expression by reducing eNOS mRNA stabilization. In the present study, we also show that the cytosol of human neutrophils contains proteins that bind to a specific region within the 3'-untranslated region (3'-UTR) of eNOS mRNA. Tumor necrosis factor-alpha increased the binding of the cytosolic proteins to the 3'-UTR of eNOS mRNA. Simvastatin reduced the TNF-alpha-related binding activity of neutrophil cytosolic proteins to eNOS mRNA, which was associated with its protective effect on eNOS protein expression. The in vivo reproduction of the in vitro findings was performed in neutrophils obtained from patients with AMI and showed a diminished expression of eNOS protein, which was associated with increased binding of the cytosolic proteins.


CONCLUSIONS
These observations demonstrate that human neutrophils express eNOS, which is downregulated by TNF-alpha and during AMI. This effect is associated with increased binding of neutrophil cytosolic proteins to the 3'-UTR of eNOS mRNA.
de Frutos
Cardiovascular Research and Hypertension Laboratory, Fundación Jiménez Díaz, Madrid, Spain.
Sánchez de Miguel
L
L
Farré
J
J
Gómez
J
J
Romero
J
J
Marcos-Alberca
P
P
Nuñez
A
A
Rico
L
L
López-Farré
A
A
D000328:Adult; D000368:Aged; D015152:Blotting, Northern; D015536:Down-Regulation / Q000502:physiology; D005260:Female; D006801:Humans; D000960:Hypolipidemic Agents / Q000494:pharmacology; D008297:Male; D008875:Middle Aged; D009203:Myocardial Infarction / Q000097:blood*; D009504:Neutrophils / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D052250:Nitric Oxide Synthase Type III; D019821:Simvastatin / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000502:physiology*
D000960:Hypolipidemic Agents; D014409:Tumor Necrosis Factor-alpha; D019821:Simvastatin; C496314:NOS3 protein, human; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III