2-2--(hydroxynitrosohydrazono)bis-ethanamine and linsidomine

Nitric oxide (NO) donors are pharmacologically active substances that release NO in vivo or in vitro. NO has a variety of functions such as the release of prostanoids, inhibition of platelet aggregation, effect on angiogenesis, and production of oxygen free radicals. This report discusses the chemical and pharmacological characteristics of NO donors, their effect on platelet function and cyclooxygenase, their cardiac action including myocardial infarction, and release of superoxide anions. This review stresses NO tolerance and the effect of NO donors on angiogenesis in myocardial infarction and in solid tumors.

Topics: Animals; Aspirin; Drug Tolerance; Heart; Humans; In Vitro Techniques; Molsidomine; Myocardial Infarction; Neovascularization, Physiologic; Nitric Oxide; Nitric Oxide Donors; Nitroglycerin; Nitroprusside; Platelet Aggregation; Prostaglandin-Endoperoxide Synthases; Superoxides; Triazenes

The biological outcome of nitric oxide (NO) and reactive nitrogen species (RNS) in regulating pro survival and pro death autophagic pathways still demand further investigation. In the present study, we investigated the effect of nitrosative stress in K562 cells using NO donor compound DETA-NONOate, peroxynitrite, and SIN-1. Exposure to NO, peroxynitrite, and SIN-1 caused decrease in K562 cell survival. NO induced autophagy but not apoptosis or necrosis in K562 cells. In contrast, peroxynitrite and SIN-1 treatment induced apoptosis in K562 cells. Surprisingly, inhibition of autophagic response using 3-methyladenine led to the induction of apoptosis in K562 cells. Increase in 5'adenosine monophosphate-activated protein kinase (AMPK) phosphorylation was only observed in the presence of NO donor indicated that AMPK was crucial to induce autophagy in K562 cells. We for the first time discovered a novel role of p73 in autophagy induction under nitrosative stress in K562 cells. TAp73α was only induced upon exposure to NO but not in the presence of peroxynitrite. Reduced glutathione (GSH)/oxidised glutathione (GSSG) ratio remained unaltered upon NO exposure. Our data suggest a complex network of interaction and cross regulations between NO and p73. These data open a new path for therapies based on the abilities of RNS to induce autophagy-mediated cell death.

Topics: Apoptosis; Autophagy; Humans; K562 Cells; Molsidomine; Nitroso Compounds; Peroxynitrous Acid; Reactive Nitrogen Species; Tumor Protein p73

Nitric oxide (NO) has emerged as an important signaling molecule in plants, but little is known about the effects of reactive nitrogen species in plant mitochondria. In this study, the effects of DETA-NONOate, a pure NO slow generator, and of SIN-1 (3-morpholinosydnonimine), a peroxynitrite producer, on the activities of respiratory pathways, enzymatic and non-enzymatic antioxidants have been investigated in isolated mitochondria from pea leaves. No significant changes in lipid peroxidation, protein oxidation or in ascorbate and glutathione redox state were observed after DETA-NONOate treatments whereas cytochrome pathway (CP) respiration was reversibly inhibited and alternative pathway (AP) respiration showed little inhibition. On the other hand, NO did not affect neither activities of Mn superoxide dismutase (Mn-SOD) nor enzymes involved in the ascorbate and glutathione regeneration in mitochondria except for ascorbate peroxidase (APX), which was reversely inhibited depending on ascorbate concentration. Finally, SIN-1 treatment of mitochondria produced a decrease in CP respiration, an increase in protein oxidation and strongly inhibited APX activity (90%), with glutathione reductase and dehydroascorbate reductase (DHAR) being moderately inhibited (30 and 20%, respectively). This treatment did not affect monodehydroascorbate reductase (MDHAR) and Mn-SOD activities. Results showed that mitochondrial nitrosative stress was not necessarily accompanied by oxidative stress. We suggest that NO-resistant AP and mitochondrial APX may be important components of the H(2) O(2) -signaling pathways under nitrosative stress induced by NO in this organelle. Also, MDHAR and DHAR, via ascorbate regeneration, could constitute an essential antioxidant defense together with Mn-SOD, against NO and ONOO(-) stress in plant mitochondria.

Topics: Antioxidants; Cell Respiration; Lipid Peroxidation; Mitochondria; Molsidomine; Nitric Oxide; Nitroso Compounds; Oxidative Stress; Pisum sativum; Plant Leaves; Reactive Nitrogen Species

Phospholipases A(2) (PLA(2)) participate in neuronal death signalling pathways because of their ability to release lipid mediators, although the contribution of each isoform and mechanism of neurotoxicity are still elusive. Using a novel fluorogenic method to assess changes in a PLA(2) activity by flow cytometry, here we show that the group IIA secretory phospholipase A(2) isoform (GIIA) was specifically activated in cortical neurons following stimulation of N-methyl-d-aspartate glutamate receptor subtype (NMDAR). For activation, GIIA required Ca(2+) and reactive oxygen/nitrogen species, and inhibition of its activity fully prevented NMDAR-mediated neuronal apoptotic death. Superoxide, nitric oxide or peroxynitrite donors stimulated GIIA activity, which mediated neuronal death. Intriguingly, we also found that GIIA activity induced mitochondrial superoxide production after NMDAR stimulation. These results reveal a novel role for GIIA in excitotoxicity both as target and producer of superoxide in a positive-loop of activation that may contribute to the propagation of neurodegeneration.

Topics: Animals; Annexin A5; Apoptosis; Calcium; Cells, Cultured; Cerebral Cortex; Dose-Response Relationship, Drug; Embryo, Mammalian; Enzyme Inhibitors; Excitatory Amino Acid Antagonists; Female; Flow Cytometry; Glutamic Acid; Group II Phospholipases A2; Mitochondria; Molsidomine; Neurons; Nitric Oxide Donors; Nitroso Compounds; Pregnancy; Rats; Rats, Wistar; Reactive Oxygen Species; Receptors, N-Methyl-D-Aspartate; Xanthine; Xanthine Oxidase

Nitric oxide (NO) is a potent nonadrenergic, noncholinergic mediator of gastrointestinal smooth muscle. We aimed to investigate the effects of new NO/cyclic guanosine monophosphate (cGMP) pathway-affecting agents at the sheep sphincter of Oddi (SO) in vitro.. Sheep SO rings were mounted in organ baths and tested for isometric tension and cGMP levels in response to 3,3-bis(aminoethyl)-1-hydroxy-2-oxo-1-triazene; 3-morpholinosydnonimine hydrochloride (SIN-1); and BAY 41-2272 in the presence and absence of 1H-(1,2,4)oxadiazole(4,3-a)quinoxalin-1-one (ODQ).. 3,3-bis(Aminoethyl)-1-hydroxy-2-oxo-1-triazene; SIN-1; and BAY 41-2272 relaxed SO rings in a concentration-dependent manner. These relaxations were significantly decreased in the presence of ODQ (P < 0.05). All agents significantly increased the cGMP levels compared with the control group (P < 0.05). The increased cGMP levels in the 3,3-bis(aminoethyl)-1-hydroxy-2-oxo-1-triazene- and BAY 41-2272-treated groups were significantly different from both control and carbachol groups (P < 0.05), whereas the increase in the SIN-1 group was significantly different from all groups (P < 0.05). The cGMP levels were significantly lower in the presence of ODQ compared with its absence (P < 0.05).. The relaxation of SO rings by these agents may be via increasing the levels of cGMP. The additional increase produced by SIN-1 may be the combined effects of NO generation and activation of guanylyl cyclase.

Topics: Animals; Carbachol; Cyclic GMP; Dose-Response Relationship, Drug; Guanylate Cyclase; In Vitro Techniques; Male; Molsidomine; Muscle Relaxation; Muscle, Smooth; Nitric Oxide; Oxadiazoles; Pyrazoles; Pyridines; Quinoxalines; Sheep; Sphincter of Oddi; Triazenes

Human leucocyte antigen G (HLA-G) is a tolerogenic molecule that protects the fetus from maternal immune attack, may favour tumoral immunoescape and is up-regulated in viral and inflammatory diseases. The aim of this work was to discover if nitric oxide (NO) could affect HLA-G expression or function because NO is an important modulator of innate and adaptive immunity. For this purpose HLA-G expression and function were analysed following treatment with a NO donor or a peroxynitrite donor in various cell lines expressing HLA-G either spontaneously or upon transfection. Results showed NO-dependent nitration of both cellular and soluble HLA-G protein, but not all HLA-G moieties underwent nitration. Endogenous biosynthesis of NO by both U-937-HLA-G1 and M8-HLA-G5 stable transfectants also caused HLA-G nitration. The NO decreased total HLA-G cellular protein content and expression on the cell surface, while increasing HLA-G shedding into the culture medium. This effect was post-transcriptional and the result of metalloprotease activity. By contrast, NO pretreatment did not affect HLA-G capability to suppress NK cytotoxicity and lymphocyte proliferation. Our studies show that NO regulates the availability of HLA-G molecules without modifying their biological activities.

Topics: Cytotoxicity, Immunologic; Dose-Response Relationship, Immunologic; Histocompatibility Antigens Class I; HLA Antigens; HLA-G Antigens; Humans; Immune Tolerance; Killer Cells, Natural; Matrix Metalloproteinases; Molsidomine; Nitric Oxide; Nitric Oxide Donors; Reverse Transcriptase Polymerase Chain Reaction; RNA Processing, Post-Transcriptional; Triazenes; Tumor Cells, Cultured

Indoleamine 2,3-dioxygenase (IDO) is an enzyme that depletes l-tryptophan, which provokes a decreased T cell response. This enzyme is expressed in human placenta, and can be also induced in many cell types such as monocytes, where endothelial (eNOS) and inducible (iNOS) nitric oxide synthases are also expressed. Previous studies have shown that nitric oxide (NO) inhibits IDO activity, which could cause a suppression of the biological function of IDO when both enzymes are coexpressed. As NO can exert different effects depending on several factors such as its concentration, we studied the effect of low concentrations of NO in the IDO activity in the U-937 and THP-1 monocytic cell lines. Results demonstrated that NO caused a bimodal effect in IDO function in IFN-gamma-stimulated monocytic cells: while high micromolar concentrations of the NO donors SIN-1 and DETA-NO decreased IDO activity, low micromolar concentrations of these NO donors increased IDO activity. Related to this, the NOS inhibitors L-NMMA and aminoguanidine, and the calmodulin antagonist W7 also decreased IDO activity. The effect of NO in IDO activity was not through cGMP production. Immunoprecipitation analysis showed a nitration of the IDO protein in unstimulated and stimulated U-937 and THP-1 cells. However, in monocyte-derived macrophages, with a higher NO production, aminoguanidine increased IDO activity, but the NOS substrate arginine decreased IDO activity. Considering the role of IDO in suppression, these results suggest a function in tolerance of the NOS enzymes depending on the NO production.

Topics: Dose-Response Relationship, Drug; Enzyme Activation; Enzyme Inhibitors; Humans; Immune Tolerance; Indoleamine-Pyrrole 2,3,-Dioxygenase; Interferon-gamma; Molsidomine; Monocytes; Nitric Oxide; Nitric Oxide Donors; Nitric Oxide Synthase Type II; Nitric Oxide Synthase Type III; omega-N-Methylarginine; Sulfonamides; Triazenes; U937 Cells

Upon Leishmania infection, macrophages are activated to produce nitrogen and oxygen radicals simultaneously. It is well established that the infected host cells rely on nitric oxide (NO) as the major weapon against the intracellular parasite. In India where leishmaniasis is endemic, the spice turmeric is used prolifically in food and for insect bites. Curcumin, the active principle of turmeric, is a scavenger of NO. This report shows that curcumin protects promastigotes and amastigotes of the visceral species, Leishmania donovani, and promastigotes of the cutaneous species, L. major, against the actions of S-nitroso-N-acetyl-D,L-penicillamine (SNAP) and DETANONOate, which release NO, 3-morpholino-sydnonimine hydrochloride (SIN-1), which releases NO and superoxide, and peroxynitrite, which is formed from the reaction of NO with superoxide. Thus, curcumin, as an antioxidant, is capable of blocking the action of both NO and NO congeners on the Leishmania parasite.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Curcumin; Leishmania donovani; Leishmania major; Mice; Mice, Inbred BALB C; Molsidomine; Nitric Oxide; Nitric Oxide Donors; Nitroso Compounds; Penicillamine; Peroxynitrous Acid

Reactive nitrogen and oxygen species (O2*-, H2O2, NO* and ONOO-) have been strongly implicated in the pathophysiology of neurodegenerative and mitochondrial diseases. In the present study, we examined the effects of nitrosative and/or nitrative stress generated by DETA-NO {(Z)-1-[2-aminoethyl-N-(2-ammonioethyl)amino]diazen-1-ium-1,2-diolate}, SIN-1 (3-morpholinosydnonimine hydrochloride) and SNP (sodium nitroprusside) on U87MG glioblastoma cybrids carrying wt (wild-type) and mutant [A3243G (Ala3243-->Gly)] mtDNA (mitochondrial genome) from a patient suffering from MELAS (mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes). The mutant cybrids had reduced activity of cytochrome c oxidase, significantly lower ATP level and decreased mitochondrial membrane potential. However, endogenous levels of reactive oxygen species were very similar in all cybrids regardless of whether they carried the mtDNA defects or not. Furthermore, the cybrids were insensitive to the nitrosative and/or nitrative stress produced by either DETA-NO or SIN-1 alone. Cytotoxicity, however, was observed in response to SNP treatment and a combination of SIN-1 and glucose-deprivation. The mutant cybrids were significantly more sensitive to these insults compared with the wt controls. Ultrastructural examination of dying cells revealed several characteristic features of autophagic cell death. We concluded that nitrosative and/or nitrative stress alone were insufficient to trigger cytotoxicity in these cells, but cell death was observed with a combination of metabolic and nitrative stress. The vulnerability of the cybrids to these types of injury correlated with the cellular energy status, which were compromised by the MELAS mutation.

Topics: Adenosine Triphosphate; Cell Death; Cell Survival; DNA, Mitochondrial; Humans; Hybrid Cells; MELAS Syndrome; Molsidomine; Mutation; Nitric Oxide Donors; Nitroprusside; Reactive Nitrogen Species; Reactive Oxygen Species; Triazenes

Nitric oxide (NO) has been reported to modulate the vascular endothelial growth factor (VEGF) gene by accumulating hypoxia-inducible factor-1alpha (HIF-1alpha) protein, but there is a contradiction among effects of various NO donors. The effects of NO donors including S-nitroso-N-acetyl-penicillamine (SNAP), S-nitroso-glutathione (GSNO), 1-hydroxy-2-oxo-3,3-bis(2-aminoethyl)-1-triazene (NOC18), 3-[(+/-)-(E)-ethyl-2(')-[(E)-hydroxyimino]-5-nitro-3-hexenecarbamoyl]-pyridine (NOR4), 3-morpholinosydnonimine (SIN-1), and nitroprusside (SNP) on the VEGF reporter gene were examined. SNAP, GSNO, NOC18, and NOR4 enhanced the VEGF reporter activity under normoxia and modulated the hypoxic induction. In contrast, SNP had only an inhibitory effect. An NO scavenger attenuated the reporter activation by NO donors except NOR4, but did not ameliorate the inhibitory effect of SNP. A reducing compound dithiothreitol suppressed NO-induced activation of the VEGF reporter gene. SNAP, GSNO, and NOC18 induced the accumulation of HIF-1alpha protein, while others did not. These results suggest that SNAP, GSNO, and NOC compounds are suitable for pharmacological studies in HIF-1-mediated VEGF gene activation by NO.

Topics: Blotting, Northern; Blotting, Western; Cell Nucleus; Dithiothreitol; Dose-Response Relationship, Drug; Endothelial Growth Factors; Genes, Reporter; Humans; Hypoxia-Inducible Factor 1, alpha Subunit; Luciferases; Lymphokines; Molsidomine; Nitric Oxide; Nitric Oxide Donors; Nitrites; Nitroprusside; Oxidation-Reduction; Penicillamine; Plasmids; Promoter Regions, Genetic; Pyridines; Reducing Agents; RNA; RNA, Messenger; S-Nitrosoglutathione; Transcription Factors; Triazenes; Tumor Cells, Cultured; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

The purpose of this study was to determine the effects of sodium nitroprusside (SNP), 2,2'-(hydroxynitrosohydrazino)bis-ethanamine (DETA/NO) and 3-morpholinosydnonimine (SIN-1), NO donors which yield different NO reactive species (NO+, NO* and peroxynitrite, respectively), as well as exogenous peroxynitrite, on gall bladder contractility. Under resting tone conditions, SNP induced a dose-dependent contraction with a maximal effect (10.3 +/- 0.7 mN, S.E.M.) at 1 mM. Consistent with these findings, SNP caused a concentration-dependent depolarization of gall bladder smooth muscle. The excitatory effects of SNP were dependent on extracellular calcium entry through L-type Ca2+ channels. Furthermore, the contraction and depolarization were sensitive to tyrosine kinase blockade, and an associated increase in tyrosine phosphorylation was detected in Western blot studies. DETA/NO induced dose-dependent relaxing effects. These relaxations were sensitive to the guanylyl cyclase inhibitor 1H-[1,2,4]oxidiazolo[4,3-a]quinoxaline-1-one (ODQ, 2 microM) but they were not altered by treatment with the potassium channel blockers tetraethylammoniun (TEA, 5 mM) and 4-aminopyridine (4-AP, 5 mM). When tested in a reducing environment (created by 2.5 mM 1,4-dithiothreitol, DTT), SNP caused a relaxation of gall bladder muscle strips. Similarly, the SNP-induced contraction was converted to a relaxation, and associated hyperpolarization, when DTT was added during the steady state of an SNP-induced response. SIN-1 (0.1 mM), which has been shown to release peroxynitrite, induced relaxing effects that were enhanced by superoxide dismutase (SOD, 50 U ml(-1)). The relaxations induced by either SIN-1 alone or SIN-1 in the presence of SOD were strengthened by catalase (1000 U ml(-1)) and abolished by ODQ pretreatment. However, exogenous peroxynitrite induced a concentration-dependent contraction, which was dependent on activation of leukotriene (LT) metabolism and extracellular calcium. The peroxynitrite-induced contraction was abolished in the presence of the peroxynitrite scavenger melatonin. These results suggest that SIN-1 behaves as an NO* rather than a peroxynitrite source. We conclude that, depending on the redox state, NO has opposing effects on the motility of the gall bladder, being a relaxing agent when in NO * form and a contracting agent when in NO+ or peroxynitrite redox species form. Knowledge of the contrasting effects of the different redox forms of NO can clarify our und

Topics: Acetylcholine; Animals; Arachidonic Acid; Atropine; Calcium; Gallbladder; Guinea Pigs; Male; Molsidomine; Muscle Contraction; Muscle, Smooth; Nitrates; Nitric Oxide; Nitric Oxide Donors; Nitroprusside; Nitroso Compounds; Oxidants; Oxidation-Reduction; Parasympatholytics; Vasodilator Agents

Leukocyte accumulation has been shown to be increased in sepsis. Moreover, in inducible nitric oxide synthase (iNOS) knockout mice, a further increase in leukocyte accumulation has been observed during sepsis, suggesting that nitric oxide (NO) may affect leukocyte/endothelial interaction. Accelerated peroxynitrite formation also occurs during sepsis. In the present study, the effect of peroxynitrite or NO on leukocyte adhesion to nitric oxide synthase (NOS)-inhibited or endotoxin-treated endothelium was examined. Bovine aortic endothelial cells were treated with either L-NAME or lipopolysaccharide (LPS) and interferon-gamma for 4 hr and subsequent leukocyte adhesion was measured. Both L-NAME and LPS treatment resulted in increased leukocyte adhesion compared with control. Neither a peroxynitrite donor, SIN-1, nor a direct NO donor, DETA-NO, had any effect on leukocyte adhesion to untreated endothelium. However, when the L-NAME or LPS-treated endothelial cells were treated simultaneously with either SIN-1 or DETA-NO, there was a significant reduction in leukocyte adhesion. Moreover, at the concentrations used in the present study, neither peroxynitrite nor NO showed harmful effects on normal cultured endothelial cells. These data demonstrating inhibition of leukocyte adhesion to endotoxin-treated endothelium suggest that peroxynitrite or NO may exert a beneficial effect during sepsis.

Topics: Animals; Cattle; Cell Adhesion; Cells, Cultured; Cyclic GMP; Endothelium, Vascular; Enzyme Inhibitors; Interferon-gamma; Leukocytes; Lipopolysaccharides; Molsidomine; NG-Nitroarginine Methyl Ester; Nitrates; Nitric Oxide; Nitric Oxide Donors; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Nitric Oxide Synthase Type III; Triazenes

A critical review of the literature indicates that the effects of nitric oxide synthase (NOS) inhibitors on focal cerebral ischemia are contradictory. In this experiment the authors methodically examined the dose-dependent effects of two NOS inhibitors and two NO donors on cortical infarction volume in an animal model of temporary focal cerebral ischemia simulating potential ischemia during neurovascular interventions.. Ninety-two Wistar rats underwent 3 hours of combined left middle cerebral artery and bilateral common carotid artery occlusion after having been anesthetized with 1% halothane. A nonselective NOS inhibitor, N(G)-nitro-L-arginine-methyl-ester (L-NAME), and two NO donors, 3-morpholinosydnonimine hydrochloride and NOC-18, DETA/NO, (Z)-1-[2(2-aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-i um-1,2-diolate, were administered intravenously 30 minutes before ischemia was induced. A selective neuronal NOS inhibitor, 7-nitroindazole (7-NI), was administered intraperitoneally in dimethyl sulfoxide (DMSO) 60 minutes before ischemia was induced. Two ischemic control groups, to which either saline or DMSO was administered, were also included in this study. Seventy-two hours after flow restoration, the animals were perfused with tetrazolium chloride for histological evaluation. Cortical infarction volume was significantly reduced by 71% in the group treated with 1 mg/kg L-NAME when compared with the saline-treated ischemic control group (27.1+/-37 mm3 compared with 92.5+/-26 mm3, p < 0.05). The NOS inhibitor 7-NI significantly reduced cortical infarction volume by 70% and by 92% at doses of 10 and 100 mg/kg: 35.2+/-32 mm3 (p < 0.05) and 9+/-13 mm3 (p < 0.005), respectively, when compared with the DMSO-treated ischemic control group (119+/-43 mm3). There was no significant difference between the saline-treated and DMSO-treated ischemic control groups. Treatment with NO donors did not significantly alter cortical infarction volume.. These results support an important role for NO in ischemic neurotoxicity and indicate that neuronal NOS inhibition may be valuable in reducing cortical injury in patients suffering temporary focal cerebral ischemia during neurovascular procedures.

Topics: Animals; Brain; Dose-Response Relationship, Drug; Enzyme Inhibitors; Indazoles; Ischemic Attack, Transient; Male; Molsidomine; NG-Nitroarginine Methyl Ester; Nitric Oxide Donors; Nitroso Compounds; Rats; Rats, Wistar

Retinal Müller glial cells express the inducible isoform (-2) of nitric oxide (NO) synthase (NOS) in vitro after stimulation by lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma) or in vivo in some retinal pathologies. Because NO may have beneficial or detrimental effects in the retina, we have used cocultures of retinal neurons with retinal Müller glial (RMG) cells from mice disrupted for the gene of NOS-2 [NOS-2 (-/-)] to clarify the role of NO in retinal neurotoxicity. We first demonstrated that NO produced by activated RMG cells was not toxic for RMG cells themselves. Second, the NO released from LPS/IFN-gamma-stimulated RMG cells induced neuronal cell death, because no neuronal cell death has been observed in cocultures with RMG cells from NOS-2 (-/-) mice and because inhibition of NOS-2 induction by transforming growth factor-beta or blockade of NO release by different NOS inhibitors prevented neuronal cell death. Addition of urate, a peroxynitrite scavenger, or superoxide dismutase partially prevented neuronal cell death induced by NO, whereas the presence of a poly(ADP-ribose) synthetase inhibitor, caspase inhibitors, or a guanylate cyclase inhibitor had no significant effect on cell death. These results demonstrated that a large release of NO from RMG cells is responsible for retinal neuronal cell death in vitro, suggesting a neurotoxic role for NO and peroxynitrite during retinal inflammatory or degenerative diseases, where RMG cells were activated.

Topics: Animals; Animals, Newborn; Cell Survival; Cells, Cultured; Coculture Techniques; Humans; Interferon-gamma; Isoenzymes; Lipopolysaccharides; Mice; Mice, Inbred C57BL; Mice, Inbred Strains; Mice, Knockout; Molsidomine; Neuroglia; Neurons; Nitric Oxide Donors; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Nitroso Compounds; omega-N-Methylarginine; Recombinant Proteins; Retina; Salmonella typhimurium; Superoxide Dismutase; Transforming Growth Factor beta

The survival of skeletal muscle myoblasts in culture after exposure either to a donor of NO, sodium nitroprusside (SNP), or ethanamine, 2,2'-(hydroxynitrosohydrazono)bis-(DETA NONOate), or to a donor of both NO and O(-)(2), 3-morpholinosydnonimine hydrochloride (SIN-1), was investigated. SIN-1 reduced clonogenic survival markedly but donors of NO alone did not. The injurious effect of SIN-1 was prevented by oxyhemoglobin or by uric acid but not by superoxide dismutase. The exposure of myoblasts to authentic peroxynitrite (ONOO(-)) or to DETA NONOate in the presence of an O(-)(2)-generating system did not reduce their survival. The results show that NO or ONOO(-) alone is not detrimental to myoblast survival and suggest that SIN-1 toxicity is, at least in part, mediated by H(2)O(2) in this myoblast culture system.

Topics: Animals; Cell Line; Cell Survival; Molsidomine; Muscle, Skeletal; Nitrates; Nitric Oxide; Nitric Oxide Donors; Nitroso Compounds; Oxyhemoglobins; Rats; Superoxide Dismutase; Superoxides; Uric Acid; Xanthine Oxidase

Protein tyrosine phosphatases (PTPs) contain an essential thiol in the active site which may be susceptible to attack by nitric oxide-derived biological oxidants. We assessed the effects of peroxynitrite, nitric oxide, and S-nitrosoglutathione on the activity of three human tyrosine phosphatases in vitro. The receptor-like T-cell tyrosine phosphatase (CD45), the non-receptor-like tyrosine phosphatase PTP1B, and leukocyte-antigen-related (LAR) phosphatase were all irreversibly inactivated by peroxynitrite in less than 1 s with IC(50) values of

Topics: Aniline Compounds; Anions; Dithiothreitol; Dose-Response Relationship, Drug; Enzyme Inhibitors; Glutathione; Humans; Leukocyte Common Antigens; Models, Molecular; Molsidomine; Nitrates; Nitric Oxide; Nitroso Compounds; Organophosphates; Organophosphorus Compounds; Protein Tyrosine Phosphatases; Receptor-Like Protein Tyrosine Phosphatases, Class 4; Receptors, Cell Surface; S-Nitrosoglutathione; Superoxide Dismutase; Tyrosine