oxypurinol and acetovanillone

Ventricular tachycardia or fibrillation (VT/VF) of focal origin due to triggered activity (TA) from delayed afterdepolarizations (DADs) is reproducibly inducible after anterior coronary artery occlusion. Both VT/VF and TA can be blocked by reducing reactive oxygen species (ROS). We tested the hypothesis that inhibition of NADPH oxidase and xanthine oxidase would block VT/VF.. 69 dogs received apocynin (APO), 4 mg/kg intraveneously (IV), oxypurinol (OXY), 4 mg/kg IV, or both APO and OXY (BOTH) agents, or saline 3 h after coronary occlusion. Endocardium from ischemic sites (3-D mapping) was sampled for Rac1 (GTP-binding protein in membrane NADPH oxidase) activation or standard microelectrode techniques. Results (mean±SE, * p<0.05): VT/VF originating from ischemic zones was blocked by APO in 6/10 *, OXY in 4/9 *, BOTH in 5/8 * or saline in 1/27; 11/16 VT/VFs blocked were focal. In isolated myocardium, TA was blocked by APO (10(-6) M) or OXY (10(-8) M). Rac1 levels in ischemic endocardium were decreased by APO or OXY.. APO and OXY suppressed focal VT/VF due to DADs, but the combination of the drugs was not more effective than either alone. Both drugs inhibited ischemic Rac1 with inhibition by OXY suggesting ROS-induced ROS. The inability to totally prevent VT/VF suggests that other mechanisms also contribute to ischemic VT.

Topics: Acetophenones; Action Potentials; Animals; Blotting, Western; Disease Models, Animal; Dogs; Female; Male; Myocardial Ischemia; NADPH Oxidases; Oxypurinol; rac1 GTP-Binding Protein; Tachycardia, Ventricular; Ultrasonography; Ventricular Fibrillation; Xanthine Oxidase

The present study is designed to investigate the role of neuronal nitric oxide synthase (nNOS) in the regulation of vascular mitogen-activated protein kinase (MAPK) activity under basal and angiotensin II (Ang II)-stimulated conditions.. Incubation with a potent nNOS inhibitor (L-VNIO) significantly increased superoxide (O2(-)) levels, with increased MAPK phosphorylation, in isolated aorta and vascular smooth muscle cells (VSMCs) from wild-type mice. Both increases were inhibited by the superoxide dismutase mimetic, tempol, but not by the peroxynitrite scavenger, FeTPPS. The levels of O2(-) and MAPK phosphorylation were higher in aorta from nNOS(-/-) mice than from wild-type mice. These parameters were suppressed by tempol and oxypurinal (a xanthine oxidase inhibitor). In isolated VSMCs or aorta from wild-type mice, Ang II stimulation markedly increased the levels of O2(-) and MAPK phosphorylation. L-VNIO significantly reduced Ang II-induced increases of these parameters. Apocynin, an NAD(P)H oxidase inhibitor, further inhibited Ang II-induced increases of these parameters compared with the L-VNIO-treated group. FeTPPS did not suppress the Ang II-induced increase of O2(-) levels, but markedly inhibited Ang II-induced MAPK phosphorylation. In contrast to the wild-type, in isolated aorta or VSMCs from nNOS(-/-) mice, Ang II failed to increase O2(-) levels and MAPK phosphorylation.. Under basal conditions, nNOS-derived NO acting as antioxidant reduces O2(-) accumulation and suppresses vascular MAPK phosphorylation. Under Ang II-stimulated conditions, NAD(P)H oxidase-derived O2(-), inducing nNOS uncoupling, potentiates the Ang II-induced increase of O2(-) generation. The generated O2(-) may react with NO to form peroxynitrite (ONOO(-)). Both O2(-) and ONOO(-) participate in Ang II-induced activation of vascular MAPK.

Topics: Acetophenones; Angiotensin II; Animals; Cells, Cultured; Cyclic N-Oxides; Male; Metalloporphyrins; Mice; Mice, Inbred C57BL; Mitogen-Activated Protein Kinases; Nitric Oxide Synthase Type I; Ornithine; Oxypurinol; Phosphorylation; Spin Labels; Superoxides; Xanthine Oxidase

This study tested the hypothesis that superoxide levels are elevated in isolated mesenteric resistance arteries (100-300 microm) from rats fed a short-term high-salt (HS) diet (4% NaCl for 3 days) compared to controls fed a low-salt (LS) diet (0.4% NaCl). Vascular relaxation induced by the superoxide dismutase mimetic tempol (4-hydroxytetramethylpiperidine-1-oxyl), the NADPH oxidase inhibitor apocynin and the xanthine/xanthine oxidase inhibitor oxypurinol was significantly larger in mesenteric arteries from animals fed HS diet compared to arteries from animals fed LS diet. Basal superoxide levels assessed via dihydroethidine (DHE) fluorescence were significantly elevated in arteries from rats fed HS diet, and were reduced by tempol, apocynin and oxypurinol, but not by L-NAME. Basal and methacholine-induced NO production (assessed by DAF-2T fluorescence) was significantly reduced in arteries from rats fed HS diet versus arteries from rats on LS diet. Impaired methacholine-induced NO release and vascular relaxation were restored by tempol and apocynin, but not by oxypurinol. These data suggest that the elevated production of superoxide by NADPH oxidase and xanthine/xanthine oxidase contribute to elevated basal superoxide levels, reduced NO release and impaired vascular relaxation in mesenteric resistance arteries of rats fed HS diet.

Topics: Acetophenones; Angiotensin II; Animals; Cyclic N-Oxides; Diet, Sodium-Restricted; Fluorescein; Fluorescent Dyes; Male; Mesenteric Arteries; Methacholine Chloride; Microscopy, Fluorescence; NADPH Oxidases; NG-Nitroarginine Methyl Ester; Nitric Oxide; Oxidative Stress; Oxypurinol; Rats; Rats, Sprague-Dawley; Sodium Chloride, Dietary; Spin Labels; Superoxides; Vascular Resistance; Vasoconstriction; Vasodilation; Xanthine; Xanthine Oxidase

The role of angiotensin II and reactive oxygen species in the exacerbation of diastolic heart failure is unknown. We examined the therapeutic effect of angiotensin blockade on hypertensive diastolic heart failure, focusing on the role of xanthine oxidoreductase and reduced nicotinamide-adenine dinucleotide phosphate oxidase, major enzymes producing reactive oxygen species. Dahl salt-sensitive hypertensive rats (DS rats) with established diastolic heart failure were given vehicle, candesartan (an angiotensin II receptor subtype 1 receptor blocker), oxypurinol (a xanthine oxidoreductase inhibitor), apocynin (a reduced nicotinamide-adenine dinucleotide phosphate oxidase inhibitor), or hydralazine (a vasodilator), and their therapeutic effects on diastolic heart failure were compared. Candesartan treatment of DS rats with established diastolic heart failure reversed cardiac remodeling, improved cardiac relaxation abnormality, and prolonged survival, being accompanied by the attenuation of the increase in cardiac superoxide, reduced nicotinamide-adenine dinucleotide phosphate oxidase, and xanthine oxidoreductase activities. Thus, the beneficial effect of candesartan in DS rats appears to be mediated by the inhibition of cardiac reactive oxygen species. Cardiac xanthine oxidoreductase inhibition with oxypurinol significantly reduced cardiac superoxide, prevented the progression of cardiac remodeling, and delayed the mortality in DS rats. Apocynin, which significantly inhibited cardiac reduced nicotinamide-adenine dinucleotide phosphate oxidase activity, prevented the exacerbation of diastolic heart failure more than hydralazine. However, compared with candesartan or oxypurinol, apocynin did not improve cardiac reactive oxygen species, remodeling, and function in DS rats. In conclusion, candesartan slowed the exacerbation of hypertensive diastolic heart failure in DS rats by causing reverse cardiac remodeling. Cardiac xanthine oxidoreductase contributed to these beneficial effects of candesartan.

Topics: Acetophenones; Angiotensin II Type 1 Receptor Blockers; Animals; Benzimidazoles; Biphenyl Compounds; Blood Pressure; Cardiac Output, Low; Heart Ventricles; Hypertension; Liver; Lung; Mitogen-Activated Protein Kinase Kinases; Myocardium; NADPH Oxidases; Oxypurinol; Rats; Rats, Inbred Dahl; Reactive Oxygen Species; Tetrazoles; Xanthine Oxidase

Endothelial nitric oxide synthase (eNOS) plays an important role in the control of myocardial oxygen consumption (MVO2) by nitric oxide (NO). A NOS isoform is present in cardiac mitochondria and it is derived from neuronal NOS (nNOS). However, the role of nNOS in the control of MVO2 remains unknown. MVO2 in left ventricular tissues from nNOS-/- mice was measured in vitro. Stimulation of NO production by bradykinin or carbachol induced a significant reduction in MVO2 in wild-type (WT) mice. In contrast to WT, bradykinin- or carbachol-induced reduction in MVO2 was attenuated in nNOS-/-. S-methyl-L-thiocitrulline, a potent isoform selective inhibitor of nNOS, had no effect on bradykinin-induced reduction in MVO2 in WT. Bradykinin-induced reduction in MVO2 in eNOS-/- mice, in which nNOS still exists, was also attenuated. The attenuated bradykinin-induced reduction in MVO2 in nNOS-/- was restored by preincubation with Tiron, ascorbic acid, Tempol, oxypurinol, or SB203850, an inhibitor of p38 kinase, but not apocynin. There was an increase in lucigenin-detectable superoxide anion (O2-) in cardiac tissues from nNOS-/- compared with WT. Tempol, oxypurinol, or SB203850 decreased O2- in all groups to levels that were not different from each other. There was an increase in phosphorylated p38 kinase normalized by total p38 kinase protein level in nNOS-/- compared with WT mice. These results indicate that a defect of nNOS increases O2- through the activation of xanthine oxidase, which is mediated by the activation of p38 kinase, and attenuates the control of MVO2 by NO derived from eNOS.

Topics: Acetophenones; Animals; Bradykinin; Carbachol; Heart; Immunoblotting; In Vitro Techniques; Mice; Mice, Inbred C57BL; Mice, Inbred Strains; Myocardium; Nerve Tissue Proteins; Nitric Oxide; Nitric Oxide Synthase; Nitric Oxide Synthase Type I; Nitric Oxide Synthase Type II; Nitric Oxide Synthase Type III; Oxygen Consumption; Oxypurinol; p38 Mitogen-Activated Protein Kinases; Penicillamine; Phosphorylation; Reactive Oxygen Species; Superoxides; Xanthine Oxidase

Endothelial dysfunction is an early manifestation of cigarette smoke (CS) toxicity. We have previously demonstrated that CS impairs nitric oxide (NO)-mediated endothelial function via increased generation of superoxide anion (O2*). In these studies, we investigated whether stable compounds present in CS activate specific pathways responsible for the increased endothelial O2* production.. Short exposure of bovine pulmonary artery endothelial cells (BPAECs), human pulmonary artery endothelial cells, and rat pulmonary arteries to CS extracts (CSEs) resulted in a large increase in O2* production (20-fold, 3-fold, and 2-fold increase, respectively; P<0.05 versus control), which was inhibited by the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitors diphenyleneiodinium, apocynin, and gp91 docking sequence-tat peptide but not by oxypurinol, the NO synthase inhibitor N(G)-nitro-L-arginine methyl ester, or the mitochondrial respiration inhibitor rotenone. Exposure of BPAECs to acrolein, a stable thiol-reactive agent found in CS, increased O2* production 5-fold, which was prevented by prior inhibition of NADPH oxidase.. These studies demonstrate that thiol-reactive stable compounds in CS can activate NADPH oxidase and increase endothelial O2* production, thereby reducing NO bioactivity and resulting in endothelial dysfunction. Clinically, these studies may contribute to the development of agents able to mitigate CS-mediated vascular toxicity.

Topics: Acetophenones; Acrolein; Animals; Cattle; Cells, Cultured; Endothelial Cells; Endothelium, Vascular; Enzyme Activation; Gene Products, tat; Humans; NADPH Oxidases; NG-Nitroarginine Methyl Ester; Nicotiana; Nitric Oxide; Onium Compounds; Oxypurinol; Pulmonary Artery; Rats; Rotenone; Smoke; Superoxides

The prevalence of the endothelium-impaired function disorders, such as hypertension and diabetes mellitus, and the severity of their complications are considerably greater in blacks than whites. Evidence has accumulated that superoxide (O2-) production and its interaction with nitric oxide (NO), yielding the strong oxidant peroxynitrite (ONOO-), play central roles in vascular pathophysiology. We hypothesized that the differences in endothelial NO/O2-/ONOO- metabolism may highlight the potential predisposition to endothelial dysfunction and cardiovascular complications prevalent in blacks.. Highly sensitive tandem electrochemical NO/O2-/ONOO- nanosensors were positioned in single human umbilical vein endothelial cells (HUVECs) isolated from blacks and whites, and the kinetics of NO/O2-/ONOO- release were recorded in vitro. HUVECs were also analyzed by Western immunoblotting and enzyme activity assays for NAD(P)H-oxidase and endothelial NO synthase (eNOS). Compared with whites, HUVECs from blacks elicited reduced release of bioactive NO with an accompanying increase in the release of both O2- and ONOO-. The greater potency of NO production because of eNOS upregulation in HUVECs from blacks is associated with a decrease in the NO bioavailability. This is due to increased NO degradation by excess O2- produced primarily by 2 enzymatic sources: NAD(P)H-oxidase and uncoupled eNOS.. Compared with whites, the steady-state NO/O2-/ONOO- balance in endothelial cells from blacks is kept closer to the redox states characteristic for the endothelium-impaired function disorders. This may explain the differences in racial predisposition to the endothelium dysfunction during ongoing vascular disturbances with the hallmark of enhanced NO inactivation within the endothelium by oxidative stress.

Topics: Acetophenones; Adult; Benzopyrans; Black or African American; Cells, Cultured; Disease Susceptibility; Endothelial Cells; Endothelium, Vascular; Female; Humans; Meclofenamic Acid; NAD; NADP; NADPH Oxidases; Nanotechnology; Nitric Oxide; Nitric Oxide Synthase; Nitric Oxide Synthase Type III; Oxidation-Reduction; Oxidative Stress; Oxypurinol; Peroxynitrous Acid; Phosphoproteins; Rotenone; Superoxides; Umbilical Veins; Vascular Diseases; White People

We previously reported that intrauterine undernutrition increased the oxidative stress by decreasing superoxide dismutase activity. In the present study, we tested whether NADPH oxidase, xanthine oxidase, cyclooxygenase or nitric oxide synthase are responsible for the increased O(2)(-) generation observed in rats submitted to intrauterine undernutrition. In addition, we investigated the effect of angiotensin II (ANG II) on O(2)(-) production via activation of NADPH oxidase.. Female pregnant Wistar rats were fed either normal or 50% of the normal intake diets, during the whole gestational period. At 16 weeks of age, the rats were used for the study of intravital fluorescence microscopy; microvascular reactivity, local ANG II concentration and AT(1), p22(phox) and gp91(phox) gene expression. In this study only the male offspring was used.. Treatment of mesenteric arterioles with the xanthine oxidase inhibitor oxypurinol, the nitric oxide synthase inhibitor L-NAME or the cyclooxygenase inhibitor diclofenac did not significantly change superoxide production. Thus, these vascular sources of superoxide were not responsible for the increased superoxide concentration. In contrast, treatment with the NADPH oxidase inhibitor apocynin significantly decreased superoxide generation and improved vascular function. On the other hand, intrauterine undernutrition did not alter the gene expression for p22(phox) and gp91(phox). The fact that the local ANG II concentration was increased and the attenuation of oxidative stress by blocking AT(1) receptor with losartan, led us to suggest that ANG II induces O(2)(-) generation in intrauterine undernourished rats.. Our study shows that NADPH oxidase inhibition attenuated superoxide anion generation and ameliorated vascular function in rats submitted to intrauterine undernutrition. Although it is not clear which mechanisms are responsible for the increase in NADPH oxidase activity, a role for ANG II-mediated superoxide production via activation of NADPH oxidase is suggested.

Topics: Acetophenones; Angiotensin II; Animals; Cyclooxygenase Inhibitors; Diclofenac; Enzyme Inhibitors; Female; Fetal Growth Retardation; Male; Mesenteric Arteries; Microscopy, Fluorescence; NADPH Oxidases; NG-Nitroarginine Methyl Ester; Nitric Oxide Synthase; Oxypurinol; Pregnancy; Random Allocation; Rats; Rats, Wistar; Renin-Angiotensin System; Superoxides; Vasodilation; Xanthine Oxidase

Accumulating evidence has suggested that cellular production of superoxide acts as an intracellular messenger to regulate gene expression and modulate cellular activities. In this report, we set out to investigate the role of active H-ras-mediated superoxide production on tumor cell malignancy in a SV-40 transformed human lung WI-38 VA-13 cell line. Stable transfection and expression of constitutively active mutant V12-H-ras (V12-H-ras) dramatically increased intracellular production of superoxide. The expression of V12-H-ras significantly enhanced cell proliferation, migration and resistance to TNF-alpha treatment compared to that of parental and vector control cells, while expression of wild type H-ras (WT-H-ras) only had modest effects. Upon scavenging by superoxide dismutase and other molecules that decrease the intracellular level of active H-ras mediated superoxide production, cell proliferation, migration and resistance to TNF-alpha were significantly reduced. Furthermore, we demonstrated that the activation of membrane NADPH oxidase activity by expression of active H-ras contributed to the intracellular superoxide production. The causal relationship between membrane superoxide production and increased cell proliferation, migration, and resistance to TNF-alpha by the expression of active H-ras, has provided direct evidence to demonstrate that superoxide acts as an intracellular messenger to cascade ras oncogenic signal relay and to modulate tumor malignant activity.

Topics: Acetophenones; Allopurinol; Cell Division; Cell Line, Transformed; Cell Movement; Drug Resistance; Enzyme Activation; Genes, ras; Genistein; Humans; Lung Neoplasms; Membrane Proteins; Models, Biological; NADPH Oxidases; Onium Compounds; Oxypurinol; Protein Kinase Inhibitors; Rotenone; Superoxides; Tumor Necrosis Factor-alpha