Measurement of Nitric Oxide and Peroxynitrite Generation in the Postischemic Heart

Wang, Penghai; Zweíer, Jay L.

doi:10.1074/jbc.271.46.29223

Cited by 521 publications

(359 citation statements)

References 51 publications

(43 reference statements)

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“…Similar values would also be expected for ONOO - [38]. In the current study, we observed that both ONOO -and O 2 .-significantly alter nNOS function at levels above 10 -20 μM while at levels above 100 μM >50% reduction in nNOS activity was seen.…”

Section: Discussionsupporting

confidence: 89%

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Dose dependent effects of reactive oxygen and nitrogen species on the function of neuronal nitric oxide synthase

Sun

Druhan

Zweíer

2008

Archives of Biochemistry and Biophysics

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Reactive nitrogen species (RNS) and oxygen species (ROS) have been reported to modulate the function of nitric oxide synthase (NOS); however, the precise dosedependent effects of specific RNS and ROS on NOS function are unknown. Questions remain unanswered regarding whether pathophysiological levels of RNS and ROS alter NOS function, and if this alteration is reversible. We measured the effects of peroxynitrite (ONOO -), superoxide (O 2 .-), hydroxyl radical ( . OH), and H 2 O 2 on nNOS activity. The results showed that NO production was inhibited in a dosedependent manner by all four oxidants, but only O 2 . -and ONOO -were inhibitory at pathophysiological concentrations (≤ 50 μM). Subsequent addition of tetrahydrobiopterin (BH 4 ) fully restored activity after O 2 .-exposure, while BH 4 partially rescued the activity decrease induced by the other three oxidants. Furthermore, treatment with either ONOO -or O 2 .-stimulated nNOS uncoupling with decreased NO and enhanced O 2 .-generation. Thus, nNOS is reversibly uncoupled by O 2 .-(≤ 50 μM), but irreversibly uncoupled and inactivated by ONOO -. Additionally, we observed that the mechanism by which oxidative stress alters nNOS activity involves not only BH 4 oxidation, but also nNOS monomerization as well as possible degradation of the heme.

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Section: Discussionsupporting

confidence: 89%

“…Nitric oxide and oxygen radical generation is increased in postischemic tissues such as the heart and brain and mediate postischemic injury [38,39]. It is well known that oxygen radicals cause lipid peroxidation and cellular calcium loading both of which are critical processes of cellular injury [34,40].…”

Section: Discussionmentioning

confidence: 99%

Dose dependent effects of reactive oxygen and nitrogen species on the function of neuronal nitric oxide synthase

Sun

Druhan

Zweíer

2008

Archives of Biochemistry and Biophysics

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“…I/RP-induced EC injury is believed to be self-inflicted, resulting from a burst of endogenous ONOO Ϫ upon the onset of RP, and, due to decreased bioavailable NO, it predisposes ECs to increased neutrophil adhesion (24,76). In agreement with that, bolus injections of ONOO Ϫ into isolated hearts were shown to acutely inhibit EC-dependent coronary vasodilation (75).…”

Section: Discussionsupporting

confidence: 60%

“…In agreement with its effects on ETC components, ONOO Ϫ potentiated the inhibition of respiration in isolated cardiac muscle compared with the inhibition elicited by NO alone and made it irreversible with time (79). Under stress conditions, such as those at reperfusion (RP) following ischemia (I), where NO production is upregulated in endothelial cells (ECs) and cardiomyocytes, either because of increases in Ca 2ϩ -dependent NOS activity via changes in shear stress and intracellular Ca 2ϩ concentration or because of expression of inducible NOS isoforms, ONOO Ϫ formation predominates over O 2 ⅐ Ϫ dismutation (24,76). ONOO Ϫ -mediated inactivation of complex I increases O 2 ⅐ Ϫ production from that location, and O 2 ⅐ Ϫ generated by complex I is released exclusively into the matrix (16,51,(63)(64)(65).…”

mentioning

confidence: 99%

Endothelial cell respiration is affected by the oxygen tension during shear exposure: role of mitochondrial peroxynitrite

Jones¹,

Han²,

Presley³

et al. 2008

American Journal of Physiology-Cell Physiology

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Cultured vascular endothelial cell (EC) exposure to steady laminar shear stress results in peroxynitrite (ONOO Ϫ ) formation intramitochondrially and inactivation of the electron transport chain. We examined whether the "hyperoxic state" of 21% O2, compared with more physiological O2 tensions (PO2), increases the shear-induced nitric oxide (NO) synthesis and mitochondrial superoxide (O2 ⅐ Ϫ ) generation leading to ONOO Ϫ formation and suppression of respiration. Electron paramagnetic resonance oximetry was used to measure O2 consumption rates of bovine aortic ECs sheared (10 dyn/cm 2 , 30 min) at 5%, 10%, or 21% O2 or left static at 5% or 21% O2. Respiration was inhibited to a greater extent when ECs were sheared at 21% O2 than at lower PO2 or left static at different PO2. Flow in the presence of an endothelial NO synthase (eNOS) inhibitor or a ONOO Ϫ scavenger abolished the inhibitory effect. EC transfection with an adenovirus that expresses manganese superoxide dismutase in mitochondria, and not a control virus, blocked the inhibitory effect. Intracellular and mitochondrial O2 ⅐ Ϫ production was higher in ECs sheared at 21% than at 5% O2, as determined by dihydroethidium and MitoSOX red fluorescence, respectively, and the latter was, at least in part, NO-dependent. Accumulation of NO metabolites in media of ECs sheared at 21% O2 was modestly increased compared with ECs sheared at lower PO2, suggesting that eNOS activity may be higher at 21% O2. Hence, the hyperoxia of in vitro EC flow studies, via increased NO and mitochondrial O2 ⅐ Ϫ production, leads to enhanced ONOO Ϫ formation intramitochondrially and suppression of respiration. shear stress; endothelium; mitochondria; reactive oxygen species MITOCHONDRIA ARE THE SUBCELLULAR organelles for the production of cellular energy, but they also activate intracellular signaling pathways that modulate cell proliferation or promote cell cycle arrest and apoptosis by oxidative or nitrosative reactions. These reactions are regulated by the matrix concentration of nitric oxide (NO), which diffuses to the mitochondria from the cytosolic NO synthase (NOS) isoforms and is thought to be produced also locally by a mitochondrial NOS variant (11). In rat heart, skeletal muscle and liver mitochondria (15,57,58), isolated rat hearts (59), and whole animals (68), NO at physiological concentrations binds reversibly and in competition with oxygen (O 2 ) to the reduced binuclear center Cu B /a 3 of cytochrome-c oxidase (complex IV) of the electron transport chain (ETC) and inhibits mitochondrial O 2 uptake. At slightly higher concentrations, NO oxidizes ubiquinol of ubiquinolcytochrome c reductase (complex III) to increase unstable ubisemiquinone, which, by univalent electron transfer to O 2 , produces the reactive O 2 species (ROS) superoxide (O 2 ⅐ Ϫ ) (57, 58). O 2 ⅐ Ϫ generated from that location is released both into the matrix and intermembrane space (51), where it is dismutated to hydrogen peroxide (H 2 O 2 ) by manganese superoxide dismutase (MnSOD) and copper zinc SOD (CuZ...

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“…Both NO and peroxynitrite, have been hypothesized to be critical mediators of postischemic injury in the heart and in other tissues. 91,92 The major pathway for the formation of NO is known to involve a class of nitric oxide synthase (NOS) enzymes, which convert arginine and oxygen to citrulline and NO. 88 NO has also been reported to be generated in biological systems by a non-enzymatic pathway, which involves chemical reduction of inorganic nitrite.…”

Section: D Mapping Of Myocardial Oxygenation In the Ischemic Heartmentioning

confidence: 99%

Cardiac applications of EPR imaging

Kuppusamy

Zweíer

2004

NMR in Biomedicine

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This review summarizes the development and application of a variety of EPR imaging modalities including spatial, spectral-spatial (spectroscopic), gated-imaging and oxygen mapping to cardiovascular studies. It has been hypothesized that free radical metabolism, oxygenation and nitric oxide generation in biological organs such as the heart may vary over the spatially defined tissue structure. We have developed instrumentation optimized for 3D spatial and 3D or 4D spectral-spatial imaging of free radicals at 1.2 GHz. Using this instrumentation high quality 3D spectral-spatial imaging of nitroxyl (nitroxide) metabolism was performed, as well as spatially localized measurements of oxygen concentrations, based on the oxygen-dependent line-broadening of the EPR spectrum. Both exogenously infused probes and endogenous radicals were used to obtain the images. It is demonstrated that the EPR imaging is a powerful tool which can provide unique information regarding the spatial localization of free radicals, oxygen and nitric oxide in biological organs and tissues.

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Measurement of Nitric Oxide and Peroxynitrite Generation in the Postischemic Heart

Abstract: Altered nitric oxide (NO ⅐)

Cited by 521 publications

References 51 publications

Dose dependent effects of reactive oxygen and nitrogen species on the function of neuronal nitric oxide synthase

Dose dependent effects of reactive oxygen and nitrogen species on the function of neuronal nitric oxide synthase

Endothelial cell respiration is affected by the oxygen tension during shear exposure: role of mitochondrial peroxynitrite

Cardiac applications of EPR imaging

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