Paradoxical fate and biological action of peroxynitrite on human platelets.

Moro, Marı́a A.; Darley‐Usmar, Victor; Goodwin, David A.; Read, Nicholas G.; Zamora-Pino, Rubén; Feelisch, Martin; Radomski, Marek W.; Moncada, Salvador

doi:10.1073/pnas.91.14.6702

Cited by 340 publications

(215 citation statements)

References 27 publications

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“…In Chinese hamster ovary cells, a higher intracellular glutathione level following treatment with a moderate dose of NAC (1 mM) led to stimulation of hyperthermiainduced cell death rather than to a protective effect (37). These and our observations are in line with reports showing that thiols, including NAC and cysteine, can form S-nitrosylated intermediates (38,39). They function as endogenous donors of nitric oxide.…”

Section: Discussionsupporting

confidence: 91%

“…The NAC-induced dissipation of Dw was also attenuated by the calcium chelator, EGTA, but not by a calcium ionophore. These treatments were used because free calcium may facilitate oxidation of NAC to a disulfide derivative (38,39). However, an increase in the dose of NAC in presence of EGTA did not prevent NAC-induced depolarization of luteal cells.…”

Section: Discussionmentioning

confidence: 99%

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N‐acetylcysteine impairs survival of luteal cells through mitochondrial dysfunction

Löhrke

Weitzel

et al. 2010

Cytometry Pt A

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N-acetylcysteine (NAC) is known as an antioxidant and used for mucus viscosity reduction. However, this drug prevents or induces cell death depending on the cell type. The response of steroidogenic luteal cells to NAC is unknown. Our data shows that NAC can behave as an antioxidant or prooxidant in dependency on the concentration and mitochondrial energization. NAC elevated the flowcytometric-measured portion of hypodiploid (dying) cells. This rise was completely abolished by aurintricarboxylic acid, an inhibitor of topoisomerase II. NAC increased the secretion of nitric oxide and cellular nitrotyrosine. An image analysis indicated that cells pretreated with NAC and loaded with DHR showed a fluorescent structure probably elicited by the oxidative product of DHR, rhodamine 123 that sequesters mitochondrially. Pretreating luteal cells with NAC or adding NAC directly to mitochondrial fractions followed by assessing the mitochondrial transmembrane potential difference (Dw) by the JC-1 technique demonstrated a marked decrease in Dw. A protonophore restored Dw and rotenone (an inhibitor of respiratory chain complex I) inhibited mitochondrial recovering. Thus, in steroidogenic luteal cells from healthy mature corpus luteum, NAC impairs cellular survival by interfering with mitochondrial metabolism. The protonophoreinduced recovering of NAC-provoked decrease in Dw indicates that an ATP synthasefavored route of H 1 re-entry to the matrix is essentially switched off by NAC while other respiratory chain complexes remain intact. These data may be important for therapeutic timing of treatments with NAC. ' 2010 International Society for Advancement of Cytometry Key terms luteal cells; antioxidant; redox imbalance; dihydrorhodamine 123; flow cytometry; mitochondrial dysfunction; viability THE corpus luteum is a transient ovarian steroidogenic gland, which develops within few days from the ovulated follicle to establish embryonic development. Intense vascularization is essential for maturation and maintenance of a healthy gland (1,2). The mature mid-luteal phase corpus luteum secretes large quantities of progesterone, a derivative from its mitochondrially formed precursor, pregnenolone.Steroidogenesis-dependent oxy-radicals and oxidants are generated during intramitochondrial conversion of cholesterol through the activity of NADPH-dependent cholesterol monooxygenase (side chain cleaving). It catalyzes in a phospholipid-assisted manner the formation of pregnenolone. This process is accompanied by an electron leakage into the mitochondrial matrix (3) and the synthesis of 4-methylpental as byproduct (4,5). In addition, regulatory constituents driving enhanced steroidogenesis are generated by the cyclooxygenase and lipoxygenase pathways which require reactive oxygen species. For instance, cyclooxygenase has an absolute requirement for hydroperoxide to catalyze oxygenation of lipids with a lipoperoxidation as the first step in forming prostanoids (6). In turn, high concentration of antioxidants and oxidant-detoxifying enzym...

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Section: Discussionsupporting

confidence: 91%

Section: Discussionmentioning

confidence: 99%

N‐acetylcysteine impairs survival of luteal cells through mitochondrial dysfunction

Löhrke

Weitzel

et al. 2010

Cytometry Pt A

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“…Also, our working hypothesis involves intermediary formation of N 2 O 3 , a potent nitrosating agent that could account for the observed peroxynitrite-induced nitrosation of GSH, especially in light of our findings that the nitrosation reaction has a pronounced pH dependence and does not occur at significant rates below pH 7.5 (45). Accordingly, reactive intermediates formed in the course decomposition to NO 2 Ϫ and O 2 could be responsible for stimulation of soluble guanylyl cyclase by peroxynitrite (45), resulting in cyclic GMP-mediated biological effects such as vascular smooth muscle relaxation and inhibition of platelet aggregation (46,47).…”

Section: Discussionmentioning

confidence: 99%

Metabolic Fate of Peroxynitrite in Aqueous Solution

Pfeiffer

Gorren

Schmidt

et al. 1997

Journal of Biological Chemistry

299

162

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Peroxynitrite, the reaction product of nitric oxide (NO) and superoxide (O 2 . ) is assumed to decompose upon protonation in a first order process via intramolecular rearrangement to NO 3 ؊ . The present study was carried out to elucidate the origin of NO 2 ؊ found in decomposed peroxynitrite solutions. As revealed by stopped-flow spectroscopy, the decay of peroxynitrite followed firstorder kinetics and exhibited a pK a of 6.8 ؎ 0.1. The reaction of peroxynitrite with NO was considered as one possible source of NO 2 ؊ , but the calculated second order rate constant of 9.1 ؋ 10 4 M ؊1 s ؊1 is probably too small to explain NO 2 ؊ formation under physiological conditions. Moreover, pure peroxynitrite decomposed to NO 2 ؊ without apparent release of NO. Determination of NO 2 ؊ and NO 3 ؊ in solutions of decomposed peroxynitrite showed that the relative amount of NO 2 ؊ increased with increasing pH, with NO 2 ؊ accounting for about 30% of decomposition products at pH 7.5 and NO 3 ؊ being the sole metabolite at pH 3.0. Formation of NO 2 ؊ was accompanied by release of stoichiometric amounts of O 2 (0.495 mol/mol of NO 2 ؊ ). The two reactions yielding NO 2 ؊ and NO 3 ؊ showed distinct temperature dependences from which a difference in E act of 26.2 ؎ 0.9 kJ mol ؊1 was calculated. The present results demonstrate that peroxynitrite decomposes with significant rates to NO 2 ؊ plus O 2 at physiological pH. Through formation of biologically active intermediates, this novel pathway of peroxynitrite decomposition may contribute to the physiology and/or cytotoxicity of NO and superoxide.The reaction between nitric oxide (NO) and superoxide anion (O 2 . ) yields peroxynitrite with a second order rate constant near the diffusion-controlled limit (k ϭ 4.3-6.7 ϫ 10 9 M Ϫ1 s Ϫ1 ) (1, 2). The reaction constitutes an important sink for O 2 . because it is about twice as fast as the maximum velocity of SOD. 1 Consequently, peroxynitrite has been implicated in many pathological conditions including stroke (3), heart disease (4), and atherosclerosis (5, 6). The potential cellular targets for peroxynitrite cytotoxicity include the antioxidants ascorbate, ␣-tocopherol, and uric acid (7-10), protein and non-protein sulfhydryls (11), DNA (12), and membrane phospholipids (13). Decomposition of peroxynitrite is complex (14, 15). The anion is rather stable in alkaline solutions but decomposes rapidly (t 1/2 ϭ 1 s at pH 7.4, 37°C) upon protonation to peroxynitrous acid (ONOOH) (pK a ϭ 6.8) (16). Two pathways of ONOOH decomposition have been proposed. Some studies have argued that ONOOH is cleaved homolytically to generate hydroxyl and NO 2 radicals. This hypothesis is based on the sensitivity to hydroxyl radical scavengers of certain peroxynitrite-induced reactions, including the formation of malondialdehyde from deoxyribose and the hydroxylation on the benzene ring of sodium benzoate, phenylalanine, and tyrosine (16, 17). Studies on decomposition of peroxynitrite by electron paramagnetic resonance spectroscopy with the spin traps 5,5-dime...

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“…Although exogenously added ONOO Ϫ causes, depending on the concentration used, apoptosis or necrosis, at the same time it is also considered a biological mediator exhibiting physiological NOrelated reactions (20,27,28). By analogy, the production of NO is not always associated with cellular toxicity.…”

Section: Discussionmentioning

confidence: 99%

Superoxide Formation and Macrophage Resistance to Nitric Oxide-mediated Apoptosis

Brüne

Götz

Meßmer

et al. 1997

Journal of Biological Chemistry

122

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RAW 264.7 macrophages, when challenged with a combination of lipopolysaccharide (10 g/ml) and interferon-␥ (100 units/ml), respond with endogenous NO ⅐ formation, which ultimately results in apoptotic cell death. Apoptosis is detected morphologically by chromatin condensation. Concomitantly we noticed the accumulation of the tumor suppressor protein p53. NO ⅐ -derived apoptosis was blocked by the NO ⅐ -synthase inhibitor N Gmonomethyl-L-arginine. Repetitive treatment of RAW 264.7 macrophages with lipopolysaccharide/interferon-␥, followed by subculturing viable cells, allowed us to select resistant macrophages which we called RES. RES cells still produced comparable amounts of nitrite/nitrate in response to agonist treatment but showed no apoptotic markers, i.e. chromatin condensation or p53 accumulation. However, RES macrophages undergo apoptosis in the presence of exogenously supplied NO ⅐ , released from the NO-donors S-nitrosoglutathione or spermine-NO. Assessment of cytochrome c reduction established that RES cells released twice the amount of superoxide compared to RAW 264.7 macrophages under both resting and stimulated conditions. We linked increased superoxide production to cellular macrophage resistance by demonstrating decreased apoptosis after simultaneous application of S-nitrosoglutathione or spermine-NO and the redox cycler 2,3-dimethoxy-1,4-naphthoquinone. Our results suggest that macrophage resistance toward NO ⅐ -mediated apoptosis is, at least in part, due to increased superoxide formation. Therefore, the balance between reactive nitrogen and reactive oxygen species regulates RAW 264.7 macrophage apoptosis.Nitric oxide (NO) 1 is recognized for its participation in diverse biological processes in nearly all aspects of life (1-3). The formation of NO ⅐ occurs under both physiological and pathophysiological settings. The molecule is synthesized by a family of enzymes termed NO ⅐ synthases (NOS), which utilize arginine as their substrate in the generation of NO ⅐ and stoichiometric amounts of citrulline (4). For convenience, two types of NOS are recognized; constitutive isoforms, which are active for a relatively short time in response to intracellular Ca 2ϩ fluctuations, and a cytokine-inducible isoform. For the latter to be active, mRNA translation and protein synthesis are required. The inducible NOS generates large amounts of NO ⅐ for an extended period. However, once NO ⅐ is produced by the action of NOS, it is extremely susceptible to both oxidation and reduction. This results in the concomitant formation of species with NO ϩ -like activity (nitrosonium ion) or NO Ϫ (nitroxyl anion), respectively (5). In addition to reacting with oxygen, superoxide, and transition metals, NO ⅐ causes biological signaling via interactions with sulfhydryl groups. Because multiple NO surrogates are formed, transduction pathways are classified as either cyclic GMP-dependent or -independent. Cyclic GMP formation is initiated by NO ⅐ binding to the heme group of soluble guanylyl cyclase, thus causing enzyme activati...

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Paradoxical fate and biological action of peroxynitrite on human platelets.

Cited by 340 publications

References 27 publications

N‐acetylcysteine impairs survival of luteal cells through mitochondrial dysfunction

N‐acetylcysteine impairs survival of luteal cells through mitochondrial dysfunction

Metabolic Fate of Peroxynitrite in Aqueous Solution

Superoxide Formation and Macrophage Resistance to Nitric Oxide-mediated Apoptosis

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