P2Y11 Agonism Prevents Hypoxia/Reoxygenation- and Angiotensin II-Induced Vascular Dysfunction and Intimal Hyperplasia Development

Vascular dysfunction in cardiovascular diseases includes vasomotor response impairments, endothelial cells (ECs) activation, and smooth muscle cells (SMCs) proliferation and migration to the intima. This results in intimal hyperplasia and vessel failure. We previously reported that activation of the P2Y11 receptor (P2Y11R) in human dendritic cells, cardiofibroblasts and cardiomyocytes was protective against hypoxia/reoxygenation (HR) lesions. In this study, we investigated the role of P2Y11R signaling in vascular dysfunction. P2Y11R activity was modulated using its pharmacological agonist NF546 and antagonist NF340. Rat aortic rings were exposed to angiotensin II (AngII) and evaluated for their vasomotor response. The P2Y11R agonist NF546 reduced AngII-induced vascular dysfunction by promoting EC-dependent vasorelaxation, through an increased nitric oxide (NO) bioavailability and reduced AngII-induced H2O2 release; these effects were prevented by the use of the P2Y11R antagonist NF340. Human vascular SMCs and ECs were subjected to AngII or H/R simulation in vitro. P2Y11R agonist modulated vasoactive factors in human ECs, that is, endothelial nitric oxide synthase (eNOS) and endothelin-1, reduced SMC proliferation and prevented the switch towards a synthetic phenotype. H/R and AngII increased ECs secretome-induced SMC proliferation, an effect prevented by P2Y11R activation. Thus, our data suggest that P2Y11R activation may protect blood vessels from HR-/AngII-induced injury and reduce vascular dysfunctions. These results open the way for new vasculoprotective interventions.

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Tyrosine nitration on calmodulin enhances calcium-dependent association and activation of nitric-oxide synthase

Journal of Biological Chemistry ◽

10.1074/jbc.ra119.010999 ◽

2019 ◽

Vol 295 (8) ◽

pp. 2203-2211 ◽

Cited By ~ 3

Author(s):

Joseph J. Porter ◽

Hyo Sang Jang ◽

Mohammad Mahfuzul Haque ◽

Dennis J. Stuehr ◽

Ryan A. Mehl

Keyword(s):

Nitric Oxide ◽

Nitric Oxide Synthase ◽

Calcium Binding ◽

Vascular Dysfunction ◽

Post Translational Modification ◽

Calcium Dependent ◽

Genetic Code Expansion ◽

Oxide Synthase ◽

Endothelial Nitric Oxide

Production of reactive oxygen species caused by dysregulated endothelial nitric-oxide synthase (eNOS) activity is linked to vascular dysfunction. eNOS is a major target protein of the primary calcium-sensing protein calmodulin. Calmodulin is often modified by the main biomarker of nitroxidative stress, 3-nitrotyrosine (nitroTyr). Despite nitroTyr being an abundant post-translational modification on calmodulin, the mechanistic role of this modification in altering calmodulin function and eNOS activation has not been investigated. Here, using genetic code expansion to site-specifically nitrate calmodulin at its two tyrosine residues, we assessed the effects of these alterations on calcium binding by calmodulin and on binding and activation of eNOS. We found that nitroTyr–calmodulin retains affinity for eNOS under resting physiological calcium concentrations. Results from in vitro eNOS assays with calmodulin nitrated at Tyr-99 revealed that this nitration reduces nitric-oxide production and increases eNOS decoupling compared with WT calmodulin. In contrast, calmodulin nitrated at Tyr-138 produced more nitric oxide and did so more efficiently than WT calmodulin. These results indicate that the nitroTyr post-translational modification, like tyrosine phosphorylation, can impact calmodulin sensitivity for calcium and reveal Tyr site-specific gain or loss of functions for calmodulin-induced eNOS activation.

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Gallic Acid Attenuates Angiotensin II-Induced Hypertension and Vascular Dysfunction by Inhibiting the Degradation of Endothelial Nitric Oxide Synthase

Frontiers in Pharmacology ◽

10.3389/fphar.2020.01121 ◽

2020 ◽

Vol 11 ◽

Cited By ~ 2

Author(s):

Xiao Yan ◽

Qi-Yu Zhang ◽

Yun-Long Zhang ◽

Xiao Han ◽

Shu-Bin Guo ◽

...

Keyword(s):

Nitric Oxide ◽

Angiotensin Ii ◽

Nitric Oxide Synthase ◽

Gallic Acid ◽

Endothelial Nitric Oxide Synthase ◽

Vascular Dysfunction ◽

Induced Hypertension ◽

Oxide Synthase ◽

Endothelial Nitric Oxide

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Ginkolide A-Gold Nanoparticles Inhibit Vascular Smooth Muscle Proliferation And Migration In Vitro And Reduces Intimal Hyperplasia In A Mouse Model

Journal of Surgical Research ◽

10.1016/j.jss.2010.11.481 ◽

2011 ◽

Vol 165 (2) ◽

pp. 253-254

Author(s):

S.M. Weakley ◽

X. Wang ◽

H. Mu ◽

P.H. Lin ◽

Q. Yao ◽

...

Keyword(s):

Gold Nanoparticles ◽

Smooth Muscle ◽

Mouse Model ◽

Vascular Smooth Muscle ◽

Intimal Hyperplasia ◽

Smooth Muscle Proliferation ◽

And Migration ◽

Proliferation And Migration

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Renal endothelial dysfunction and impaired autoregulation after ischemia-reperfusion injury result from excess nitric oxide

AJP Renal Physiology ◽

10.1152/ajprenal.00302.2005 ◽

2006 ◽

Vol 291 (3) ◽

pp. F619-F628 ◽

Cited By ~ 32

Author(s):

Zhengrong Guan ◽

Glenda Gobé ◽

Desley Willgoss ◽

Zoltán H. Endre

Keyword(s):

Nitric Oxide ◽

Renal Failure ◽

Angiotensin Ii ◽

Endothelial Dysfunction ◽

Ischemia Reperfusion Injury ◽

Ischemia Reperfusion ◽

Artery Occlusion ◽

Tubuloglomerular Feedback ◽

Sprague Dawley ◽

Endothelial Nitric Oxide

Endothelial dysfunction in ischemic acute renal failure (IARF) has been attributed to both direct endothelial injury and to altered endothelial nitric oxide synthase (eNOS) activity, with either maximal upregulation of eNOS or inhibition of eNOS by excess nitric oxide (NO) derived from iNOS. We investigated renal endothelial dysfunction in kidneys from Sprague-Dawley rats by assessing autoregulation and endothelium-dependent vasorelaxation 24 h after unilateral (U) or bilateral (B) renal artery occlusion for 30 (U30, B30) or 60 min (U60, B60) and in sham-operated controls. Although renal failure was induced in all degrees of ischemia, neither endothelial dysfunction nor altered facilitation of autoregulation by 75 pM angiotensin II was detected in U30, U60, or B30 kidneys. Baseline and angiotensin II-facilitated autoregulation were impaired, methacholine EC50 was increased, and endothelium-derived hyperpolarizing factor (EDHF) activity was preserved in B60 kidneys. Increasing angiotensin II concentration restored autoregulation and increased renal vascular resistance (RVR) in B60 kidneys; this facilitated autoregulation, and the increase in RVR was abolished by 100 μM furosemide. Autoregulation was enhanced by Nω-nitro-l-arginine methyl ester. Peri-ischemic inhibition of inducible NOS ameliorated renal failure but did not prevent endothelial dysfunction or impaired autoregulation. There was no significant structural injury to the afferent arterioles with ischemia. These results suggest that tubuloglomerular feedback is preserved in IARF but that excess NO and probably EDHF produce endothelial dysfunction and antagonize autoregulation. The threshold for injury-producing, detectable endothelial dysfunction was higher than for the loss of glomerular filtration rate. Arteriolar endothelial dysfunction after prolonged IARF is predominantly functional rather than structural.

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Endothelium-specific sepiapterin reductase deficiency in DOCA-salt hypertension

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00835.2011 ◽

2012 ◽

Vol 302 (11) ◽

pp. H2243-H2249 ◽

Cited By ~ 20

Author(s):

Ji Youn Youn ◽

Ting Wang ◽

John Blair ◽

Karine M. Laude ◽

Jeong-Ho Oak ◽

...

Keyword(s):

Nitric Oxide ◽

Angiotensin Ii ◽

Oxidase Inhibitor ◽

Aortic Endothelial Cells ◽

Sepiapterin Reductase ◽

Enos Uncoupling ◽

Induced Hypertension ◽

Salt Hypertension ◽

Endothelial Nitric Oxide ◽

Aortic Endothelial

The endothelial nitric oxide synthase (eNOS) requires tetrahydrobiopterin (H4B) as a cofactor and, in its absence, produces superoxide (O2·−) rather than nitric oxide (NO·), a condition referred to as eNOS uncoupling. DOCA-salt-induced hypertension is associated with H4B oxidation and uncoupling of eNOS. The present study investigated whether administration of sepiapterin or H4B recouples eNOS in DOCA-salt hypertension. Bioavailable NO· detected by electron spin resonance was markedly reduced in aortas of DOCA-salt hypertensive mice. Preincubation with sepiapterin (10 μmol/l for 30 min) failed to improve NO· bioavailability in hypertensive aortas while it augmented NO· production from control vessels, implicating a hypertension-associated deficiency in sepiapterin reductase (SPR), the rate-limiting enzyme for sepiapterin conversion to H4B. Indeed, a decreased SPR expression was observed in aortic endothelial cells, but not in endothelium-denuded aortic remains, implicating an endothelium-specific SPR deficiency. Administration of hypertensive aortas with H4B (10 μmol/l, 30 min) partially restored vascular NO· production. Combined administration of H4B and the NADPH oxidase inhibitor apocynin (100 μmol/l, 30 min) fully restored NO· bioavailability while reducing O2·− production. In angiotensin II-induced hypertension, however, aortic endothelial SPR expression was not affected. In summary, administration of sepiapterin is not effective in recoupling eNOS in DOCA-salt hypertension, due to an endothelium-specific loss in SPR, whereas coadministration of H4B and apocynin is highly efficient in recoupling eNOS. This is consistent with our previous observations that in angiotensin II hypertension, endothelial deficiency in dihydrofolate reductase is alternatively responsible for uncoupling of eNOS. Taken together, these data indicate that strategies specifically targeting at different H4B metabolic enzymes might be necessary in restoring eNOS function in different types of hypertension.

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Inflammatory Monocytes Determine Endothelial Nitric-oxide Synthase Uncoupling and Nitro-oxidative Stress Induced by Angiotensin II

Journal of Biological Chemistry ◽

10.1074/jbc.m114.604231 ◽

2014 ◽

Vol 289 (40) ◽

pp. 27540-27550 ◽

Cited By ~ 70

Author(s):

Sabine Kossmann ◽

Hanhan Hu ◽

Sebastian Steven ◽

Tanja Schönfelder ◽

Daniela Fraccarollo ◽

...

Keyword(s):

Oxidative Stress ◽

Nitric Oxide ◽

Angiotensin Ii ◽

Nitric Oxide Synthase ◽

Endothelial Nitric Oxide Synthase ◽

Inflammatory Monocytes ◽

Oxide Synthase ◽

Endothelial Nitric Oxide

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In vivo gene transfer of endothelial nitric oxide synthase to carotid arteries from hypercholesterolemic rabbits enhances endothelium-dependent relaxations but does not reverse vascular dysfunction

Atherosclerosis ◽

10.1016/s0021-9150(99)80224-x ◽

1999 ◽

Vol 144 ◽

pp. 58

Author(s):

T. O'Brien ◽

J. Sato ◽

T. Mohasci ◽

A. Noel ◽

P. Gloviczki ◽

...

Keyword(s):

Nitric Oxide ◽

Nitric Oxide Synthase ◽

Gene Transfer ◽

Endothelial Nitric Oxide Synthase ◽

Carotid Arteries ◽

Vascular Dysfunction ◽

Oxide Synthase ◽

Endothelial Nitric Oxide ◽

In Vivo Gene Transfer

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Alpha-Asarone Protects Endothelial Cells from Injury by Angiotensin II

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2014/682041 ◽

2014 ◽

Vol 2014 ◽

pp. 1-7 ◽

Cited By ~ 3

Author(s):

Hai-Xia Shi ◽

Jiajun Yang ◽

Tao Yang ◽

Yong-Liang Xue ◽

Jun Liu ◽

...

Keyword(s):

Nitric Oxide ◽

Endothelial Cells ◽

Angiotensin Ii ◽

Cell Injury ◽

Fluorescent Dyes ◽

Umbilical Vein ◽

Protective Effects ◽

Ang Ii ◽

Versus Model

α-Asarone is the major therapeutical constituent ofAcorus tatarinowiiSchott. In this study, the potential protective effects ofα-asarone against endothelial cell injury induced by angiotensin II were investigatedin vitro. The EA.hy926 cell line derived from human umbilical vein endothelial cells was pretreated withα-asarone (10, 50, 100 µmol/L) for 1 h, followed by coincubation with Ang II (0.1 µmol/L) for 24 h. Intracellular nitric oxide (NO) and reactive oxygen species (ROS) were detected by fluorescent dyes, and phosphorylation of endothelial nitric oxide synthase (eNOS) atSer1177was determined by Western blotting.α-Asarone dose-dependently mitigated the Ang II-induced intracellular NO reduction (P<0.01versus model) and ROS production (P<0.01versus model). Furthermore, eNOS phosphorylation (Ser1177) by acetylcholine was significantly inhibited by Ang II, while pretreatment for 1 h withα-asarone partially prevented this effect (P<0.05versus model). Additionally, cell viability determined by the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay (105~114.5% versus control,P>0.05) was not affected after 24 h of incubation withα-asarone at 1–100 µmol/L. Therefore,α-asarone protects against Ang II-mediated damage of endothelial cells and may be developed to prevent injury to cardiovascular tissues.

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