Role of N-acetyl-N-nitroso-tryptophan as nitric oxide donor in the modulation of HIF-1-dependent signaling

2010 ◽  
Vol 391 (5) ◽  
pp. 533-540 ◽  
Author(s):  
Utta Berchner-Pfannschmidt ◽  
Suzan Tug ◽  
Jun Hu ◽  
Buena Delos Reyes ◽  
Joachim Fandrey ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Mrna Expression ◽  
Animal Studies ◽  
Hypoxia Inducible Factor ◽  
Nitric Oxide Donor ◽  
Strong Increase ◽  
No Donor ◽  
No Donors ◽  
Hypoxic Conditions

Abstract N-Acetyl-N-nitroso-tryptophan (NANT) is well known for its capacity to generate nitric oxide (NO)-releasing compounds. It is unknown, however, whether NANT can be successfully applied as a precursor of NO in a complex biological environment such as a cell culture system. NO donors can be useful to induce the transcription factor hypoxia-inducible factor 1 (HIF-1) that coordinates the protection of cells and tissues from the lack of oxygen, termed hypoxia. HIF-1 degradation is controlled by prolyl hydroxylase 2 (PHD2) which needs to be inhibited for HIF-1 accumulation. Here, the effects of NANT in inhibiting recombinant PHD2 and up-regulating of HIF-1 and HIF-1-mediated carboanhydrase-9 (CA9) mRNA expression were compared in living cells with the NO donors N-nitrosomelatonin (NOMela) and S-nitrosoglutathione (GSNO) under normoxic and hypoxic conditions. In contrast to GSNO, NANT was similar to NOMela being highly effective in inhibiting recombinant PHD2. NANT-mediated activation of HIF-1 in oxygenated cells was comparable to hypoxic activation of HIF-1 in all cases. In contrast, under hypoxia NANT was able to boost hypoxic cellular HIF-1 levels by further reducing the activity of cellular PHD2. The strong increase of HIF-dependent CA9 mRNA expression demonstrated that NANT-induced HIF-1 was transcriptionally active. Finally, the efficacy of NANT to increase both HIF-1 and CA9 mRNA did not depend on the absolute conformation of the tryptophan moiety. In conclusion, NANT appears to be an excellent NO donor for cells in culture and l-NANT should be useful for in vivo animal studies.

Increased endothelin-1 and endothelin receptor expression in myocytes of ischemic and reperfused rat hearts and ventricular myocytes exposed to ischemic conditions and its inhibition by nitric oxide generation

2003 ◽  
Vol 81 (2) ◽  
pp. 105-113 ◽  
Author(s):  
Xiaohong Tracey Gan ◽  
Subrata Chakrabarti ◽  
Morris Karmazyn
Keyword(s):  
Nitric Oxide ◽  
Mrna Expression ◽  
Ventricular Myocytes ◽  
Receptor Expression ◽  
Neonatal Rat ◽  
No Donor ◽  
Myocardial Ischemia And Reperfusion ◽  
Endothelin 1 ◽  
Hypoxic Conditions ◽  
Rat Hearts

Endothelin-1 (ET-1) and nitric oxide (NO) exert opposite effects in the cardiovascular system, and there is evidence that the NO counters the potential deleterious effects of ET-1. We investigated whether NO affects the increased mRNA expression of ET-1 and endothelin receptors induced by (i) 30 min of ischemia with or without 30 min reperfusion in myocytes from isolated rat hearts or (ii) ischemic conditions (acidosis or hypoxia) in cultured rat neonatal ventricular myocytes. Ischemia with or without reperfusion produced more than a twofold increase in mRNA expression of ET-1 as well as the ETAand ETBreceptor (P < 0.05), although these effects were completely blocked by the NO donor 3-morpholinosydnonimine (SIN-1; 1 μM). To assess the possible factors regulating ET expression, myocytes were exposed to acidosis (pH 6.8–6.2) or to hypoxic conditions in an anaerobic chamber for 24 h in the presence or absence of SIN-1. At all acidic pHs, ET-1 and ETAreceptor mRNA expression was significantly (P < 0.05) elevated approximately threefold, although the magnitude of elevation was independent of the degree of acidosis. These effects were completely prevented by SIN-1. ETBreceptor expression was unaffected by acidosis. Hypoxia increased ET-1 as well as ETAand ETBreceptor expression threefold (P < 0.05), although this was unaffected by SIN-1. Our results demonstrate that myocardial ischemia and reperfusion upregulate the ET system, which is inhibited by NO. Although increased expression of the ET system can be mimicked by both acidosis and hypoxia, only the effects of the former are NO sensitive. NO may serve an endogenous inhibitory factor which regulates the expression of the ET system under pathological conditions.Key words: ET-1, ET receptors, NO, neonatal rat ventricular myocytes, hypoxia, acidosis.

Hypoxia enhances a cGMP-independent nitric oxide relaxing mechanism in pulmonary arteries

2003 ◽  
Vol 285 (2) ◽  
pp. L296-L304 ◽  
Author(s):  
Christopher J. Mingone ◽  
Sachin A. Gupte ◽  
Takafumi Iesaki ◽  
Michael S. Wolin
Keyword(s):  
Nitric Oxide ◽  
Sarcoplasmic Reticulum ◽  
Myosin Light Chain ◽  
Soluble Guanylate Cyclase ◽  
Pulmonary Arteries ◽  
Cyclopiazonic Acid ◽  
No Donor ◽  
No Donors ◽  
Hypoxic Conditions ◽  
Guanylate Cyclase Activation

Nitric oxide (NO) donors generally relax vascular preparations through cGMP-mediated mechanisms. Relaxation of endothelium-denuded bovine pulmonary arteries (BPA) and coronary arteries to the NO donor S-nitroso- N-acetyl-penicillamine (SNAP) is almost eliminated by inhibition of soluble guanylate cyclase activation with 10 μM 1H-[1,2,4]oxadiazolo-[4,3-a]quinoxalin-1-one (ODQ), whereas only a modest inhibition of relaxation is observed under hypoxia (PO2 = 8–10 Torr). This effect of hypoxia is independent of the contractile agent used and is also observed with NO gas. ODQ eliminated SNAP-induced increases in cGMP under hypoxia in BPA. cGMP-independent relaxation of BPA to SNAP was not attenuated by inhibition of K+ channels (10 mM tetraethylammonium), myosin light chain phosphatase (0.5 μM microcystin-LR), or adenylate cyclase (4 μM 2′,5′-dideoxyadenosine). SNAP relaxed BPA contracted with serotonin under Ca2+-free conditions in the presence of hypoxia and ODQ, and contraction to Ca2+ readdition was also attenuated. The sarcoplasmic reticulum Ca2+-reuptake inhibitor cyclopiazonic acid (0.2 mM) attenuated SNAP-mediated relaxation of BPA in the presence of ODQ. Thus hypoxic conditions appear to promote a cGMP-independent relaxation of BPA to NO by enhancing sarcoplasmic reticulum Ca2+ reuptake.

scholarly journals Sildenafil-nitric oxide donor combination promotes ventricular tachyarrhythmias in the swine right ventricle

2002 ◽  
Vol 282 (5) ◽  
pp. H1787-H1792 ◽  
Author(s):  
Moshe Swissa ◽  
Toshihiko Ohara ◽  
Moon-Hyoung Lee ◽  
Sanjay Kaul ◽  
Prediman K. Shah ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Donor ◽  
Tyrode Solution ◽  
No Donor ◽  
Maximum Slope ◽  
No Donors ◽  
Restitution Curve ◽  
Rapid Pacing ◽  
Drug Free ◽  
Endocardial Activation

We tested the hypothesis that sildenafil, singly or in combination with nitric oxide (NO) donors, promotes ventricular tachycardia (VT) and ventricular fibrillation (VF). Vulnerability to VT/VF was tested by rapid pacing in eight isolated normal swine right ventricles (RV). The endocardial activation was optically mapped, and the dynamic action potential duration (APD) restitution curves were constructed with metal microelectrodes. At baseline, no VT/VF could be induced. Sildenafil (0.2 μg/ml) or NO donor singly or in combination did not alter VT/VF vulnerability. However, when 2 μg/ml sildenafil was combined with NO donors, the incidence of VT and VF rose significantly ( P < 0.01). VT with a single periodic wavefront was induced in five of eight RVs, and VF with multiple wavefronts was induced in all eight RVs. The sildenafil-NO donor pro-VT/VF combination significantly increased the maximum slope of the APD restitution curve and the amplitude of the APD alternans. The pro-VT/VF effects of sildenafil were reversible after drug-free Tyrode solution perfusion. We conclude that a sildenafil (2 μg/ml) and NO donor combination increases VT/VF vulnerability in the normal RV by a mechanism compatible with the restitution hypothesis.

Effect of in vivo nitrate tolerance on hypersensitivity to NO donors after NO-synthase blockade

2002 ◽  
Vol 80 (11) ◽  
pp. 1106-1118 ◽  
Author(s):  
Jodan D Ratz ◽  
Michael A Adams ◽  
Brian M Bennett
Keyword(s):  
Nitric Oxide ◽  
Glyceryl Trinitrate ◽  
Nitrate Tolerance ◽  
Metabolite Formation ◽  
No Donor ◽  
No Donors ◽  
Nos Inhibitors ◽  
Pressure Lowering ◽  
Oxide Synthase

Animals treated with nitric oxide synthase (NOS) inhibitors exhibit marked hypersensitivity to the blood pressure lowering effects of exogenous nitric oxide (NO) donors. We used this model as a sensitive index to evaluate the relative importance of reduced biotransformation of glyceryl trinitrate (GTN) to NO in the development of nitrate tolerance. NOS-blockade hypertension using NG-nitro-L-arginine methyl ester (L-NAME) caused a marked enhancement of the mean arterial pressure (MAP) decrease mediated by GTN in nontolerant rats. However, even large doses of GTN were unable to change the MAP in GTN-tolerant, NOS-blockade hypertensive animals. In contrast, the MAP responses to the spontaneous NO donor sodium nitroprusside (SNP) were completely unaltered in either tolerant rats or tolerant NOS-blockade hypertensive animals, indicating that NO-dependent vasodilatory mechanisms remain intact despite the development of GTN tolerance. The MAP-lowering effects of GTN in NOS-blockade hypertensive animals were restored 48 h after cessation of chronic GTN exposure. These alterations in the pharmacodynamic response to GTN during tolerance development and reversal were associated with parallel changes in the pattern of GTN metabolite formation, suggesting that the activity of one or more enzymes involved in nitrate metabolism was altered as a consequence of chronic GTN exposure. These findings suggest that the vasodilation resulting from the vascular biotransformation of GTN to NO (or a closely related species) is severely compromised in nitrate-tolerant animals, and that although other mechanisms may contribute to the vascular changes observed following the development of GTN tolerance, decreased GTN bioactivation is likely the most important.Key words: biotransformation, glyceryl trinitrate, hypertension, nitric oxide, tolerance.

scholarly journals Involvement of nitric oxide donor compounds in the bactericidal activity of human neutrophils in vitro

2003 ◽  
Vol 52 (4) ◽  
pp. 303-308 ◽  
Author(s):  
Magdalena Klink ◽  
Maciej Cedzyński ◽  
Anna St Świerzko ◽  
Henryk Tchórzewski ◽  
Zofia Sulowska
Keyword(s):  
Nitric Oxide ◽  
Coronary Heart Disease ◽  
Heart Disease ◽  
Bactericidal Activity ◽  
Nitric Oxide Donor ◽  
Human Neutrophils ◽  
Bacterial Strains ◽  
No Donor ◽  
No Donors

The bactericidal activity of human neutrophils against extracellular and facultatively intracellular bacteria was studied in the presence of the nitric oxide (NO) donors sodium nitroprusside (SNP) and 3-morpholinosydnonimine (SIN-1), a molsidomine metabolite. SNP and molsidomine are drugs commonly used as nitrovasodilators in coronary heart disease. It is demonstrated here that the NO donor compounds themselves did not affect the viability and survival of the bacterial strains tested. Neither SNP nor SIN-1 had any effect on the process of bacteria ingestion. In contrast, NO donors enhanced the ability of neutrophils to kill Escherichia coli, Proteus vulgaris and Salmonella Anatum. However, strains differed in their susceptibility to SNP- and SIN-1-mediated killing by neutrophils. Removal of the superoxide anion reduced the bactericidal activity of SNP- and SIN-1-treated neutrophils against E. coli and S. Anatum. This suggests that the NO derivatives formed in the reaction of NO generated from donors with the reactive oxygen species released by phagocytosed neutrophils potentiate the bactericidal activity of human neutrophils in vitro. The above original observation discussed here suggests clinical significance for the treatment of patients with nitrovasodilators in the course of coronary heart disease therapy.

Effects of the nitric oxide donor 3-morpholinosydnonimine (SIN-1) in focal cerebral ischemia dependent on intracellular brain pH

2002 ◽  
Vol 97 (4) ◽  
pp. 914-921 ◽  
Author(s):  
Bernard A. Coert ◽  
Robert E. Anderson ◽  
Fredric B. Meyer
Keyword(s):  
Nitric Oxide ◽  
Cerebral Ischemia ◽  
Focal Cerebral Ischemia ◽  
Nitric Oxide Donor ◽  
Protective Agent ◽  
No Donor ◽  
Content Type ◽  
Infarction Volume ◽  
Fine Print

Object. A nitric oxide (NO) donor that has been successfully used in the treatment of myocardial infarction, 3-morpholinosydnonimine (SIN-1), may be a potential neuroprotective agent. Production of NO in brain microsomes is dependent on the pH. The purpose of this study was to determine the efficacy of SIN-1 and its dependence on pH in vivo during periods of focal cerebral ischemia. Methods. At 0.1 or 1 mg/kg, SIN-1 was administered to 54 Wistar rats 30 minutes before a 2-hour period of focal cerebral ischemia under moderate hypo-, normo-, and hyperglycemic conditions. Measurements of brain intracellular pH (pHi); regional cortical blood flow, and the redox state of nicotinamide adenine dinucleotide were obtained in three additional animals to confirm the effects of the serum glucose manipulations. The animals were killed at 72 hours after the ischemic period to obtain infarction volumes. Administration of SIN-1 significantly reduced infarction in normoglycemic animals and, to a lesser extent, in hyperglycemic animals, indicating that SIN-1 was less effective under hyperglycemic conditions. At either dose SIN-1 had no significant effect on infarction volume in moderately hypoglycemic animals because moderate hypoglycemia in itself significantly (p < 0.005) reduced infarction volume. Conclusions. The NO donor SIN-1 may be a useful intraoperative cerebral protective agent. Furthermore, it is hypothesized that a mechanism that could explain the published discrepancies regarding the effects of NO donors in vivo may be affected by differences in ischemic brain acidosis.

Hemolytic Rate and Nitric Oxide Bioavailability Modulate the Expression of Hypoxia-Inducible Factor-1α (HIF-1α) in Transgenic Sickle Mice

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 2501-2501
Author(s):  
Dhananjay K. Kaul ◽  
Trisha Dasgupta ◽  
Xiaoqin Zhang ◽  
Mary Fabry
Keyword(s):  
Oxidative Stress ◽  
Nitric Oxide ◽  
Hypoxia Inducible Factor ◽  
Control Level ◽  
Tissue Hypoxia ◽  
Heme Oxygenase 1 ◽  
Hypoxia Inducible Factor 1Α ◽  
Hypoxic Conditions ◽  
No Bioavailability

Abstract In sickle cell disease (SCD), tissue hypoxia (caused by vaso-occlusive events, abnormal red cell rheology and anemia), intense oxidative stress and persistent hemolysis are three major components that determine pathogenesis. In SCD, ischemic events are likely to activate hypoxia-inducible factor-1α (HIF-1α), a transcription factor that is involved in regulation of several genes including genes for vasoactive molecules. Although tissue ischemia is the primary instigator of HIF-1α activation, the induction, stabilization and degradation of HIF-1α also involves participation of signaling molecules, oxygen sensors (prolyl hydroxylases or PHDs) and redox reactions. Nitric oxide (NO) is a major signaling molecule affecting the activity of PHDs. We hypothesize that increased oxidative stress and reduced NO bioavailability caused by cell-free plasma heme will have a significant effect on HIF-1α expression under in vivo conditions. To this end, we have explored the effect of hemolysis and NO bioavailability on HIF-1α expression: in transgenic-knockout sickle (BERK) mice expressing varying levels of anti-sickling fetal hemoglobin (HbF), and in transgenic sickle (NY1DD) mice subjected to arginine treatment and hypoxia. We used BERK mice expressing different levels of HbF: BERK (HbF &lt;1.0), BERKγM (~20% HbF) and BERKγH (~40% HbF). BERK mice show severe pathology, tissue hypoxia, significant hemolysis, and accumulation of HIF-1α under ambient conditions as determined in the cremaster tissue. Also, BERK mice showed maximal hemolytic rate (measured as plasma heme), and introduction of γ-transgene to elevate HbF levels to 20 and 40% caused significant reductions in plasma heme values as reported (Kaul et al. JCI, 2004). Increasing HbF expression and reducing hemolysis in BERK mice resulted in corresponding increases in NO metabolites (NOx) levels (P&lt;0.05, multiple comparisons by ANOVA). Notably, in BERK mice, HIF-1α expression decreased by almost 50% with increased NOx levels. The present in vivo finding is in contrast with the reported NO-provoked HIF-1α accumulation in vitro under normoxic conditions. Next, we evaluated the effect of NO bioavailability on HIF-1α accumulation under hypoxic conditions. To this end, we treated in C57BL and transgenic sickle (NY1DD) mice with arginine (5% arginine in mouse chow). NY1DD mice show mild pathology but develop severe phenotype under hypoxic conditions. Both groups of mice were treated for 15 days with arginine followed by hypoxia (8% O2) starting day 12 of arginine treatment. Compared with normoxic controls, hypoxia caused marked accumulation of HIF-1α in the cremaster tissue. Importantly, arginine markedly reduced hypoxia-induced HIF-1α activation in NY1DD mice to the control level, indicating that under hypoxic conditions increased NO bioavailability (arginine treatment) is associated with degradation of HIF-1α. Interestingly, arginine-treated NY1DD mice show increased NOx levels, and reduced expression of heme oxygenase-1 (HO-1), a marker of hemolysis, by almost 40%. Taken together, these results provide the first in vivo demonstration that the hemolytic rate and NO bioavailability have a major influence on HIF-1α expression in the mouse models of SCD.

P076. In vivo metabolic profiling and target-tissue biochemical variegation of nitric oxide (NO) donors: Nitrite and NO as determinants of NO-donor biology

Nitric Oxide ◽  
2006 ◽  
Vol 14 (4) ◽  
pp. 41
Author(s):  
Nathan S. Bryan ◽  
David R. Janero ◽  
Fumito Saijo ◽  
Michael E. Augustyniak ◽  
Vijay Dhawan ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Metabolic Profiling ◽  
Target Tissue ◽  
No Donor ◽  
No Donors

scholarly journals The contribution of N2O3 to the cytotoxicity of the nitric oxide donor DETA/NO: an emerging role for S-nitrosylation

2013 ◽  
Vol 33 (2) ◽  
Author(s):  
Ahlam A. Ali ◽  
Jonathan A. Coulter ◽  
Claire H. Ogle ◽  
Marie M. Migaud ◽  
David G. Hirst ◽  
...  
Keyword(s):  
Oxygen Tension ◽  
Hypoxia Inducible Factor ◽  
Tumour Microenvironment ◽  
Nitric Oxide Donor ◽  
Apoptotic Pathway ◽  
No Donor ◽  
Hypoxic Conditions ◽  
Fluorimetric Analysis ◽  
First Time ◽  
The Relationship

The relationship between the biological activity of NO and its chemistry is complex. The objectives of this study were to investigate the influence of oxygen tension on the cytotoxicity of the NO• donor DETA/NO and to determine the effects of oxygen tension on the key RNS (reactive nitrogen species) responsible for any subsequent toxicity. The findings presented in this study indicate that the DETA/NO-mediated cytotoxic effects were enhanced under hypoxic conditions. Further investigations revealed that neither ONOO− (peroxynitrite) nor nitroxyl was generated. Fluorimetric analysis in the presence of scavengers suggest for the first time that another RNS, dinitrogen trioxide may be responsible for the cytotoxicity with DETA/NO. Results showed destabilization of HIF (hypoxia inducible factor)-1α and depletion of GSH levels following the treatment with DETA/NO under hypoxia, which renders cells more susceptible to DETA/NO cytotoxicity, and could account for another mechanism of DETA/NO cytotoxicity under hypoxia. In addition, there was significant accumulation of nuclear p53, which showed that p53 itself might be a target for S-nitrosylation following the treatment with DETA/NO. Both the intrinsic apoptotic pathway and the Fas extrinsic apoptotic pathway were also activated. Finally, GAPDH (glyceraldehyde-3-phosphate dehydrogenase) is another important S-nitrosylated protein that may possibly play a key role in DETA/NO-mediated apoptosis and cytotoxicity. Therefore this study elucidates further mechanisms of DETA/NO mediated cytotoxicity with respect to S-nitrosylation that is emerging as a key player in the signalling and detection of DETA/NO-modified proteins in the tumour microenvironment.

Effect of a Nitric Oxide Donor (Glyceryl Trinitrate) on Nociceptive Thresholds in Man

Cephalalgia ◽  
1996 ◽  
Vol 16 (3) ◽  
pp. 169-174 ◽  
Author(s):  
LL Thomsen ◽  
J Brennum ◽  
HK Iverson ◽  
J Olesen
Keyword(s):  
Nitric Oxide ◽  
Glyceryl Trinitrate ◽  
Animal Studies ◽  
Crossover Design ◽  
Nitric Oxide Donor ◽  
Temporal Region ◽  
Double Blind ◽  
No Donor ◽  
Pain Detection ◽  
Nociceptive Input

Several animal studies suggest that nitric oxide (NO) plays a role in central and peripheral modulation of nociception. Glyceryl trinitrate GTN) exerts its physiological actions via donation of NO. The purpose of the present study was to examine the effect of this NO donor on nociceptive thresholds in man. On two different study days separated by at least, week 12 healthy subjects received a staircase infusion of GTN (0.015, 0.25. 1.0, 2.0 mg/kg/min. 20 min each dose) or placebo in a randomized double-blind crossover design. Before the infusion and after 15 min of infusion on each dose, pressure pain detection and tolerance thresholds were determined by pressure a gometry (Somomedic AB, Sweden) in three different anatomic regions (finger, a temporal region with interposed myofascial tissue and a temporal region without interposed myofascial tissue. Relative to placebo, the three higher GTN doses induced a decrease in both detection and tolerance thresholds in the temporal region with interposed myofascial tissue ( p=0.003 detection and p=0.002 tolerance threshold: Friedman). No such changes were observed in the other two stimulated regions. These results could reflect central facilitation of nociception by NO. However, we regard convergence, of nociceptive input from pericranial myofascial tissue and from cephalic blood vessels dilated by NO as a more likely, explanation of our findings.

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