Characterization of neutral endopeptidase in vascular cells, modulation of vasoactive peptide levels.
We characterized neutral endopeptidase activity and protein in the three aortic layers and in corresponding cultured primary cells. Neutral endopeptidase was expressed in all three layers of rat aorta with higher protein level and activity in the adventitia than in the media and intimal endothelium. Neutral endopeptidase was also found in primary cultured fibroblasts, smooth muscle and endothelial cells derived from the corresponding layers. Neutral endopeptidase activity and protein were higher in the fibroblasts and smooth muscle cells than in endothelial cells. Neutral endopeptidase inhibition prevented atrial natriuretic peptide (ANP) degradation in endothelial and smooth muscle cells. It potentiated ANP-stimulated cyclic GMP production in these cells. Neutral endopeptidase inhibition also reduced bradykinin degradation and potentiated bradykinin-stimulated release of arachidonic acid in fibroblasts and endothelial cells. Our data demonstrate the presence and functional activity of neutral endopeptidase in all three cell layers of rat aorta as well as in primary cells of the vessel. The data suggest that local concentrations of vasoactive peptides in the vessel wall might be regulated by the neutral endopeptidase cleavage pathway in the immediate vicinity of their target cells.
González
W
W
U460 Institut National de la Santé et de la Recherche Médicale, Faculté X, Bichat, Paris, France.
Soleilhac
J M
JM
Fournié-Zaluski
M C
MC
Roques
B P
BP
Michel
J B
JB
D000818:Animals; D001013:Aorta, Thoracic / Q000166:cytology / Q000201:enzymology / Q000502:physiology; D016718:Arachidonic Acid / Q000378:metabolism; D009320:Atrial Natriuretic Factor / Q000378:metabolism / Q000494:pharmacology; D001920:Bradykinin / Q000378:metabolism / Q000494:pharmacology; D002469:Cell Separation; D006152:Cyclic GMP / Q000096:biosynthesis; D004591:Electrophoresis, Polyacrylamide Gel; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000201:enzymology* / Q000502:physiology*; D015260:Neprilysin / Q000737:chemistry / Q000378:metabolism*; D010455:Peptides / Q000502:physiology*; D016133:Polymerase Chain Reaction; D012313:RNA / Q000032:analysis / Q000302:isolation & purification; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction / Q000187:drug effects
D010455:Peptides; D016718:Arachidonic Acid; D012313:RNA; D009320:Atrial Natriuretic Factor; D015260:Neprilysin; D006152:Cyclic GMP; D001920:Bradykinin