Glutathionylation mediates angiotensin II-induced eNOS uncoupling, amplifying NADPH oxidase-dependent endothelial dysfunction

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Glutathionylation mediates angiotensin II-induced eNOS uncoupling, amplifying NADPH oxidase-dependent endothelial dysfunction. / Galougahi, Keyvan Karimi; Liu, Chia-Chi; Gentile, Carmine; Kok, Cindy; Nunez, Andrea; Garcia, Alvaro; Fry, Natasha A S; Davies, Michael Jonathan; Hawkins, Clare Louise; Rasmussen, Helge H; Figtree, Gemma A.

In: American Heart Association. Journal. Cardiovascular and Cerebrovascular Disease, Vol. 3, No. 2, 2014, p. e000731.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Galougahi, KK, Liu, C-C, Gentile, C, Kok, C, Nunez, A, Garcia, A, Fry, NAS, Davies, MJ, Hawkins, CL, Rasmussen, HH & Figtree, GA 2014, 'Glutathionylation mediates angiotensin II-induced eNOS uncoupling, amplifying NADPH oxidase-dependent endothelial dysfunction', American Heart Association. Journal. Cardiovascular and Cerebrovascular Disease, vol. 3, no. 2, pp. e000731. https://doi.org/10.1161/JAHA.113.000731

APA

Galougahi, K. K., Liu, C-C., Gentile, C., Kok, C., Nunez, A., Garcia, A., ... Figtree, G. A. (2014). Glutathionylation mediates angiotensin II-induced eNOS uncoupling, amplifying NADPH oxidase-dependent endothelial dysfunction. American Heart Association. Journal. Cardiovascular and Cerebrovascular Disease, 3(2), e000731. https://doi.org/10.1161/JAHA.113.000731

Vancouver

Galougahi KK, Liu C-C, Gentile C, Kok C, Nunez A, Garcia A et al. Glutathionylation mediates angiotensin II-induced eNOS uncoupling, amplifying NADPH oxidase-dependent endothelial dysfunction. American Heart Association. Journal. Cardiovascular and Cerebrovascular Disease. 2014;3(2):e000731. https://doi.org/10.1161/JAHA.113.000731

Author

Galougahi, Keyvan Karimi ; Liu, Chia-Chi ; Gentile, Carmine ; Kok, Cindy ; Nunez, Andrea ; Garcia, Alvaro ; Fry, Natasha A S ; Davies, Michael Jonathan ; Hawkins, Clare Louise ; Rasmussen, Helge H ; Figtree, Gemma A. / Glutathionylation mediates angiotensin II-induced eNOS uncoupling, amplifying NADPH oxidase-dependent endothelial dysfunction. In: American Heart Association. Journal. Cardiovascular and Cerebrovascular Disease. 2014 ; Vol. 3, No. 2. pp. e000731.

Bibtex

@article{14acf9c24a0c420fbbacae0397246626,
title = "Glutathionylation mediates angiotensin II-induced eNOS uncoupling, amplifying NADPH oxidase-dependent endothelial dysfunction",
abstract = "BACKGROUND: Glutathionylation of endothelial nitric oxide synthase (eNOS) {"}uncouples{"} the enzyme, switching its function from nitric oxide (NO) to O2(•-) generation. We examined whether this reversible redox modification plays a role in angiotensin II (Ang II)-induced endothelial dysfunction.METHODS AND RESULTS: Ang II increased eNOS glutathionylation in cultured human umbilical vein endothelial cells (HUVECs), rabbit aorta, and human arteries in vitro. This was associated with decreased NO bioavailability and eNOS activity as well as increased O2(•-) generation. Ang II-induced decrease in eNOS activity was mediated by glutathionylation, as shown by restoration of function by glutaredoxin-1. Moreover, Ang II-induced increase in O2(•-) and decrease in NO were abolished in HUVECs transiently transfected, with mutant eNOS rendered resistant to glutathionylation. Ang II effects were nicotinamide adenine dinucleotide phosphate (NADPH) oxidase dependent because preincubation with gp 91ds-tat, an inhibitor of NADPH oxidase, abolished the increase in eNOS glutathionylation and loss of eNOS activity. Functional significance of glutathionylation in intact vessels was supported by Ang II-induced impairment of endothelium-dependent vasorelaxation that was abolished by the disulfide reducing agent, dithiothreitol. Furthermore, attenuation of Ang II signaling in vivo by administration of an angiotensin converting enzyme (ACE) inhibitor reduced eNOS glutathionylation, increased NO, diminished O2(•-), improved endothelium-dependent vasorelaxation and reduced blood pressure.CONCLUSIONS: Uncoupling of eNOS by glutathionylation is a key mediator of Ang II-induced endothelial dysfunction, and its reversal is a mechanism for cardiovascular protection by ACE inhibition. We suggest that Ang II-induced O2(•-) generation in endothelial cells, although dependent on NADPH oxidase, is amplified by glutathionylation-dependent eNOS uncoupling.",
keywords = "Angiotensin II, Animals, Aorta, Blood Pressure, Cells, Cultured, Endothelial Cells, Endothelium, Vascular, Enzyme Inhibitors, Glutathione, Human Umbilical Vein Endothelial Cells, Humans, Male, Mutation, NADPH Oxidase, Nitric Oxide, Nitric Oxide Synthase Type III, Oxidation-Reduction, Rabbits, Signal Transduction, Superoxides, Transfection, Vasodilation",
author = "Galougahi, {Keyvan Karimi} and Chia-Chi Liu and Carmine Gentile and Cindy Kok and Andrea Nunez and Alvaro Garcia and Fry, {Natasha A S} and Davies, {Michael Jonathan} and Hawkins, {Clare Louise} and Rasmussen, {Helge H} and Figtree, {Gemma A}",
year = "2014",
doi = "10.1161/JAHA.113.000731",
language = "English",
volume = "3",
pages = "e000731",
journal = "American Heart Association. Journal. Cardiovascular and Cerebrovascular Disease",
issn = "2047-9980",
publisher = "Wiley-Blackwell",
number = "2",

}

RIS

TY - JOUR

T1 - Glutathionylation mediates angiotensin II-induced eNOS uncoupling, amplifying NADPH oxidase-dependent endothelial dysfunction

AU - Galougahi, Keyvan Karimi

AU - Liu, Chia-Chi

AU - Gentile, Carmine

AU - Kok, Cindy

AU - Nunez, Andrea

AU - Garcia, Alvaro

AU - Fry, Natasha A S

AU - Davies, Michael Jonathan

AU - Hawkins, Clare Louise

AU - Rasmussen, Helge H

AU - Figtree, Gemma A

PY - 2014

Y1 - 2014

N2 - BACKGROUND: Glutathionylation of endothelial nitric oxide synthase (eNOS) "uncouples" the enzyme, switching its function from nitric oxide (NO) to O2(•-) generation. We examined whether this reversible redox modification plays a role in angiotensin II (Ang II)-induced endothelial dysfunction.METHODS AND RESULTS: Ang II increased eNOS glutathionylation in cultured human umbilical vein endothelial cells (HUVECs), rabbit aorta, and human arteries in vitro. This was associated with decreased NO bioavailability and eNOS activity as well as increased O2(•-) generation. Ang II-induced decrease in eNOS activity was mediated by glutathionylation, as shown by restoration of function by glutaredoxin-1. Moreover, Ang II-induced increase in O2(•-) and decrease in NO were abolished in HUVECs transiently transfected, with mutant eNOS rendered resistant to glutathionylation. Ang II effects were nicotinamide adenine dinucleotide phosphate (NADPH) oxidase dependent because preincubation with gp 91ds-tat, an inhibitor of NADPH oxidase, abolished the increase in eNOS glutathionylation and loss of eNOS activity. Functional significance of glutathionylation in intact vessels was supported by Ang II-induced impairment of endothelium-dependent vasorelaxation that was abolished by the disulfide reducing agent, dithiothreitol. Furthermore, attenuation of Ang II signaling in vivo by administration of an angiotensin converting enzyme (ACE) inhibitor reduced eNOS glutathionylation, increased NO, diminished O2(•-), improved endothelium-dependent vasorelaxation and reduced blood pressure.CONCLUSIONS: Uncoupling of eNOS by glutathionylation is a key mediator of Ang II-induced endothelial dysfunction, and its reversal is a mechanism for cardiovascular protection by ACE inhibition. We suggest that Ang II-induced O2(•-) generation in endothelial cells, although dependent on NADPH oxidase, is amplified by glutathionylation-dependent eNOS uncoupling.

AB - BACKGROUND: Glutathionylation of endothelial nitric oxide synthase (eNOS) "uncouples" the enzyme, switching its function from nitric oxide (NO) to O2(•-) generation. We examined whether this reversible redox modification plays a role in angiotensin II (Ang II)-induced endothelial dysfunction.METHODS AND RESULTS: Ang II increased eNOS glutathionylation in cultured human umbilical vein endothelial cells (HUVECs), rabbit aorta, and human arteries in vitro. This was associated with decreased NO bioavailability and eNOS activity as well as increased O2(•-) generation. Ang II-induced decrease in eNOS activity was mediated by glutathionylation, as shown by restoration of function by glutaredoxin-1. Moreover, Ang II-induced increase in O2(•-) and decrease in NO were abolished in HUVECs transiently transfected, with mutant eNOS rendered resistant to glutathionylation. Ang II effects were nicotinamide adenine dinucleotide phosphate (NADPH) oxidase dependent because preincubation with gp 91ds-tat, an inhibitor of NADPH oxidase, abolished the increase in eNOS glutathionylation and loss of eNOS activity. Functional significance of glutathionylation in intact vessels was supported by Ang II-induced impairment of endothelium-dependent vasorelaxation that was abolished by the disulfide reducing agent, dithiothreitol. Furthermore, attenuation of Ang II signaling in vivo by administration of an angiotensin converting enzyme (ACE) inhibitor reduced eNOS glutathionylation, increased NO, diminished O2(•-), improved endothelium-dependent vasorelaxation and reduced blood pressure.CONCLUSIONS: Uncoupling of eNOS by glutathionylation is a key mediator of Ang II-induced endothelial dysfunction, and its reversal is a mechanism for cardiovascular protection by ACE inhibition. We suggest that Ang II-induced O2(•-) generation in endothelial cells, although dependent on NADPH oxidase, is amplified by glutathionylation-dependent eNOS uncoupling.

KW - Angiotensin II

KW - Animals

KW - Aorta

KW - Blood Pressure

KW - Cells, Cultured

KW - Endothelial Cells

KW - Endothelium, Vascular

KW - Enzyme Inhibitors

KW - Glutathione

KW - Human Umbilical Vein Endothelial Cells

KW - Humans

KW - Male

KW - Mutation

KW - NADPH Oxidase

KW - Nitric Oxide

KW - Nitric Oxide Synthase Type III

KW - Oxidation-Reduction

KW - Rabbits

KW - Signal Transduction

KW - Superoxides

KW - Transfection

KW - Vasodilation

U2 - 10.1161/JAHA.113.000731

DO - 10.1161/JAHA.113.000731

M3 - Journal article

C2 - 24755153

VL - 3

SP - e000731

JO - American Heart Association. Journal. Cardiovascular and Cerebrovascular Disease

JF - American Heart Association. Journal. Cardiovascular and Cerebrovascular Disease

SN - 2047-9980

IS - 2

ER -

ID: 128973602