20-Hydroxyeicosatetraenoic acid causes endothelial dysfunction via eNOS uncoupling

Jennifer Cheng, Jing Song Ou, Harpreet Singh, J R Falck, Dubasi Narsimhaswamy, Kirkwood A. Pritchard, Michal Laniado Schwartzman

Research output: Contribution to journalArticle

109 Citations (Scopus)

Abstract

Nitric oxide (NO), generated from L-arginine by endothelial nitric oxide synthase (eNOS), is a key endothelial-derived factor whose bioavailability is essential to the normal function of the endothelium. Endothelium dysfunction is characterized by loss of NO bioavailability because of either reduced formation or accelerated degradation of NO. We have recently reported that overexpression of vascular cytochrome P-450 (CYP) 4A in rats caused hypertension and endothelial dysfunction driven by increased production of 20- hydroxyeicosatetraenoic acid (20-HETE), a major vasoconstrictor eicosanoid in the microcirculation. To further explore cellular mechanisms underlying CYP4A-20-HETE-driven endothelial dysfunction, the interactions between 20-HETE and the eNOS-NO system were examined in vitro. Addition of 20-HETE to endothelial cells at concentrations as low as 1 nM reduced calcium ionophore-stimulated NO release by 50%. This reduction was associated with a significant increase in superoxide production. The increase in superoxide in response to 20-HETE was prevented by NG-nitro-L-arginine methyl ester, suggesting that uncoupled eNOS is a source of this superoxide. The response to 20-HETE was specific in that 19-HETE did not affect NO or superoxide production, and, in fact, the response to 20-HETE could be competitively antagonized by 19(R)-HETE. 20-HETE had no effect on phosphorylation of eNOS protein at serine-1179 or threonine-497 following addition of calcium ionophore; however, 20-HETE inhibited association of eNOS with 90-kDa heat shock protein (HSP90). In vivo, impaired acetylcholine-induced relaxation in arteries overexpressing CYP4A was associated with a marked reduction in the levels of phosphorylated vasodilator-stimulated phosphoprotein, an indicator of bioactive NO, that was reversed by inhibition of 20-HETE synthesis or action. Because association of HSP90 with eNOS is critical for eNOS activation and coupled enzyme activity, inhibition of this association by 20-HETE may underlie the mechanism, at least in part, by which increased CYP4A expression and activity cause endothelial dysfunction.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Heart and Circulatory Physiology
Volume294
Issue number2
DOIs
StatePublished - Feb 2008

Fingerprint

Nitric Oxide Synthase Type III
Cytochrome P-450 CYP4A
Nitric Oxide
Superoxides
HSP90 Heat-Shock Proteins
Calcium Ionophores
Biological Availability
Endothelium
20-hydroxy-5,8,11,14-eicosatetraenoic acid
Enzyme Activation
Eicosanoids
NG-Nitroarginine Methyl Ester
Vasoconstrictor Agents
Threonine
Microcirculation
Serine
Acetylcholine
Blood Vessels
Arginine
Endothelial Cells

Keywords

  • Cytochrome P-450
  • Endothelial nitric oxide synthase
  • Nitric oxide
  • Superoxide
  • Vascular endothelium
  • Vasodilator-stimulated phosphoprotein

ASJC Scopus subject areas

  • Physiology

Cite this

20-Hydroxyeicosatetraenoic acid causes endothelial dysfunction via eNOS uncoupling. / Cheng, Jennifer; Ou, Jing Song; Singh, Harpreet; Falck, J R; Narsimhaswamy, Dubasi; Pritchard, Kirkwood A.; Schwartzman, Michal Laniado.

In: American Journal of Physiology - Heart and Circulatory Physiology, Vol. 294, No. 2, 02.2008.

Research output: Contribution to journalArticle

Cheng, Jennifer ; Ou, Jing Song ; Singh, Harpreet ; Falck, J R ; Narsimhaswamy, Dubasi ; Pritchard, Kirkwood A. ; Schwartzman, Michal Laniado. / 20-Hydroxyeicosatetraenoic acid causes endothelial dysfunction via eNOS uncoupling. In: American Journal of Physiology - Heart and Circulatory Physiology. 2008 ; Vol. 294, No. 2.
@article{ca6c85a9b1fb4a64889bd33c71dee9b3,
title = "20-Hydroxyeicosatetraenoic acid causes endothelial dysfunction via eNOS uncoupling",
abstract = "Nitric oxide (NO), generated from L-arginine by endothelial nitric oxide synthase (eNOS), is a key endothelial-derived factor whose bioavailability is essential to the normal function of the endothelium. Endothelium dysfunction is characterized by loss of NO bioavailability because of either reduced formation or accelerated degradation of NO. We have recently reported that overexpression of vascular cytochrome P-450 (CYP) 4A in rats caused hypertension and endothelial dysfunction driven by increased production of 20- hydroxyeicosatetraenoic acid (20-HETE), a major vasoconstrictor eicosanoid in the microcirculation. To further explore cellular mechanisms underlying CYP4A-20-HETE-driven endothelial dysfunction, the interactions between 20-HETE and the eNOS-NO system were examined in vitro. Addition of 20-HETE to endothelial cells at concentrations as low as 1 nM reduced calcium ionophore-stimulated NO release by 50{\%}. This reduction was associated with a significant increase in superoxide production. The increase in superoxide in response to 20-HETE was prevented by NG-nitro-L-arginine methyl ester, suggesting that uncoupled eNOS is a source of this superoxide. The response to 20-HETE was specific in that 19-HETE did not affect NO or superoxide production, and, in fact, the response to 20-HETE could be competitively antagonized by 19(R)-HETE. 20-HETE had no effect on phosphorylation of eNOS protein at serine-1179 or threonine-497 following addition of calcium ionophore; however, 20-HETE inhibited association of eNOS with 90-kDa heat shock protein (HSP90). In vivo, impaired acetylcholine-induced relaxation in arteries overexpressing CYP4A was associated with a marked reduction in the levels of phosphorylated vasodilator-stimulated phosphoprotein, an indicator of bioactive NO, that was reversed by inhibition of 20-HETE synthesis or action. Because association of HSP90 with eNOS is critical for eNOS activation and coupled enzyme activity, inhibition of this association by 20-HETE may underlie the mechanism, at least in part, by which increased CYP4A expression and activity cause endothelial dysfunction.",
keywords = "Cytochrome P-450, Endothelial nitric oxide synthase, Nitric oxide, Superoxide, Vascular endothelium, Vasodilator-stimulated phosphoprotein",
author = "Jennifer Cheng and Ou, {Jing Song} and Harpreet Singh and Falck, {J R} and Dubasi Narsimhaswamy and Pritchard, {Kirkwood A.} and Schwartzman, {Michal Laniado}",
year = "2008",
month = "2",
doi = "10.1152/ajpheart.01172.2007",
language = "English (US)",
volume = "294",
journal = "American Journal of Physiology - Heart and Circulatory Physiology",
issn = "0363-6135",
publisher = "American Physiological Society",
number = "2",

}

TY - JOUR

T1 - 20-Hydroxyeicosatetraenoic acid causes endothelial dysfunction via eNOS uncoupling

AU - Cheng, Jennifer

AU - Ou, Jing Song

AU - Singh, Harpreet

AU - Falck, J R

AU - Narsimhaswamy, Dubasi

AU - Pritchard, Kirkwood A.

AU - Schwartzman, Michal Laniado

PY - 2008/2

Y1 - 2008/2

N2 - Nitric oxide (NO), generated from L-arginine by endothelial nitric oxide synthase (eNOS), is a key endothelial-derived factor whose bioavailability is essential to the normal function of the endothelium. Endothelium dysfunction is characterized by loss of NO bioavailability because of either reduced formation or accelerated degradation of NO. We have recently reported that overexpression of vascular cytochrome P-450 (CYP) 4A in rats caused hypertension and endothelial dysfunction driven by increased production of 20- hydroxyeicosatetraenoic acid (20-HETE), a major vasoconstrictor eicosanoid in the microcirculation. To further explore cellular mechanisms underlying CYP4A-20-HETE-driven endothelial dysfunction, the interactions between 20-HETE and the eNOS-NO system were examined in vitro. Addition of 20-HETE to endothelial cells at concentrations as low as 1 nM reduced calcium ionophore-stimulated NO release by 50%. This reduction was associated with a significant increase in superoxide production. The increase in superoxide in response to 20-HETE was prevented by NG-nitro-L-arginine methyl ester, suggesting that uncoupled eNOS is a source of this superoxide. The response to 20-HETE was specific in that 19-HETE did not affect NO or superoxide production, and, in fact, the response to 20-HETE could be competitively antagonized by 19(R)-HETE. 20-HETE had no effect on phosphorylation of eNOS protein at serine-1179 or threonine-497 following addition of calcium ionophore; however, 20-HETE inhibited association of eNOS with 90-kDa heat shock protein (HSP90). In vivo, impaired acetylcholine-induced relaxation in arteries overexpressing CYP4A was associated with a marked reduction in the levels of phosphorylated vasodilator-stimulated phosphoprotein, an indicator of bioactive NO, that was reversed by inhibition of 20-HETE synthesis or action. Because association of HSP90 with eNOS is critical for eNOS activation and coupled enzyme activity, inhibition of this association by 20-HETE may underlie the mechanism, at least in part, by which increased CYP4A expression and activity cause endothelial dysfunction.

AB - Nitric oxide (NO), generated from L-arginine by endothelial nitric oxide synthase (eNOS), is a key endothelial-derived factor whose bioavailability is essential to the normal function of the endothelium. Endothelium dysfunction is characterized by loss of NO bioavailability because of either reduced formation or accelerated degradation of NO. We have recently reported that overexpression of vascular cytochrome P-450 (CYP) 4A in rats caused hypertension and endothelial dysfunction driven by increased production of 20- hydroxyeicosatetraenoic acid (20-HETE), a major vasoconstrictor eicosanoid in the microcirculation. To further explore cellular mechanisms underlying CYP4A-20-HETE-driven endothelial dysfunction, the interactions between 20-HETE and the eNOS-NO system were examined in vitro. Addition of 20-HETE to endothelial cells at concentrations as low as 1 nM reduced calcium ionophore-stimulated NO release by 50%. This reduction was associated with a significant increase in superoxide production. The increase in superoxide in response to 20-HETE was prevented by NG-nitro-L-arginine methyl ester, suggesting that uncoupled eNOS is a source of this superoxide. The response to 20-HETE was specific in that 19-HETE did not affect NO or superoxide production, and, in fact, the response to 20-HETE could be competitively antagonized by 19(R)-HETE. 20-HETE had no effect on phosphorylation of eNOS protein at serine-1179 or threonine-497 following addition of calcium ionophore; however, 20-HETE inhibited association of eNOS with 90-kDa heat shock protein (HSP90). In vivo, impaired acetylcholine-induced relaxation in arteries overexpressing CYP4A was associated with a marked reduction in the levels of phosphorylated vasodilator-stimulated phosphoprotein, an indicator of bioactive NO, that was reversed by inhibition of 20-HETE synthesis or action. Because association of HSP90 with eNOS is critical for eNOS activation and coupled enzyme activity, inhibition of this association by 20-HETE may underlie the mechanism, at least in part, by which increased CYP4A expression and activity cause endothelial dysfunction.

KW - Cytochrome P-450

KW - Endothelial nitric oxide synthase

KW - Nitric oxide

KW - Superoxide

KW - Vascular endothelium

KW - Vasodilator-stimulated phosphoprotein

UR - http://www.scopus.com/inward/record.url?scp=39149110293&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=39149110293&partnerID=8YFLogxK

U2 - 10.1152/ajpheart.01172.2007

DO - 10.1152/ajpheart.01172.2007

M3 - Article

C2 - 18156192

AN - SCOPUS:39149110293

VL - 294

JO - American Journal of Physiology - Heart and Circulatory Physiology

JF - American Journal of Physiology - Heart and Circulatory Physiology

SN - 0363-6135

IS - 2

ER -