Endothelin-1 induces NAD(P)H oxidase in human endothelial cells.
Superoxide anions (O(*-)(2)) induce oxidative stress and reduce endothelial NO availability by peroxynitrite formation. In human endothelial cells gp91(phox) was identified as the limiting subunit of the forming NAD(P)H oxidase. Because endothelin-1 (ET-1) is considered as a pro-atherosclerotic stimulus, we analyzed the effect of ET-1 on gp91(phox) expression and O(*-)(2) generation in primary cultures of human umbilical vein endothelial cells (HUVECs). The gp91(phox) mRNA expression was quantified by standard calibrated competitive reverse transcriptase-polymerase chain reaction. ET-1 induces gp91(phox) mRNA expression in HUVEC (max. after 1 h). The induction of gp91(phox) expression was dose-dependent, reaching its maximum at 10 nmol/L ET-1. The increased gp91(phox) expression is mediated by endothelial receptor type B (ET(B)). Furthermore, ET-1 augments O(*-)(2) generation in human endothelial cells as measured by coelenterazine chemiluminescence. These data support a new mechanism: how ET-1 increases oxidative stress in the vessel wall leading to endothelial dysfunction and enhanced susceptibility to atherosclerosis.
Duerrschmidt
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N
Institute of Pathophysiology, Martin Luther University Halle-Wittenberg, Magdeburger Strasse 18, Halle, D-06097, Germany.
Wippich
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N
Goettsch
W
W
Broemme
H J
HJ
Morawietz
H
H
D002478:Cells, Cultured; D065128:Endothelin Receptor Antagonists; D019332:Endothelin-1 / Q000494:pharmacology*; D004730:Endothelium, Vascular / Q000187:drug effects / Q000502:physiology*; D005786:Gene Expression Regulation / Q000187:drug effects / Q000502:physiology; D006801:Humans; D007700:Kinetics; D008562:Membrane Glycoproteins / Q000235:genetics*; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases / Q000235:genetics*; D009842:Oligopeptides / Q000494:pharmacology; D010880:Piperidines / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics; D044023:Receptor, Endothelin B; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013481:Superoxides / Q000378:metabolism; D014158:Transcription, Genetic; D014471:Umbilical Veins
D065128:Endothelin Receptor Antagonists; D019332:Endothelin-1; D008562:Membrane Glycoproteins; D009842:Oligopeptides; D010880:Piperidines; D012333:RNA, Messenger; D044023:Receptor, Endothelin B; D013481:Superoxides; C086539:BQ 788; C056558:CYBB protein, human; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases