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 journal › Journal article › Research › peer-review
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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 - Journal of the American Heart Association
JF - Journal of the American Heart Association
SN - 2047-9980
IS - 2
ER -
ID: 128973602