Electron paramagnetic resonance evidence that cellular oxygen toxicity is caused by the generation of superoxide and hydroxyl free radicals
Open accessAbstract
Cells require molecular oxygen for the generation of energy through mitochondrial oxidative phosphorylation; however, high concentrations of oxygen are toxic and can cause cell death. A number of different mechanisms have been proposed to cause cellular oxygen toxicity. One hypothesis is that reactive oxygen free radicals may be generated; however free radical generation in hyperoxic cells has never been directly measured and the mechanism of this radical generation is unknown. In order to determine if cellular oxygen toxicity is free radical mediated, we applied electron paramagnetic resonance, EPR, spectroscopy using the spin trap 5,5′‐dimethyl‐1‐pyrroline‐N‐oxide, DMPO, to measure free radical generation in hyperoxic pulmonary endothelial cells. Cells in air did not give rise to any detectable signal. However, cells exposed to 100% O 2 for 30 min exhibited a prominent signal of trapped hydroxyl radical, DMPO‐OH, while cell free buffer did not give rise to any detectable radical generation. This cellular radical generation was demonstrated to be derived from the superoxide radical since the observed signal was totally quenched by superoxide dismutase, but not by equal concentrations of the denatured enzyme. It was confirmed that the hydroxyl radical was generated since in the presence of ethanol the CH 3·CH(OH) radical was formed. Loss of cell viability as measured by uptake of trypan blue dye was observed paralleling the measured free radical generation. Thus, superoxide and hydroxyl radicals are generated in hyperoxic pulmonary endothelial cells and this appears to be an important mechanism of cellular oxygen toxicity.
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Mitochondrial uncoupling and increased ROS generation activates the mitochondrial permeability transition pore (mPTP), allowing the release of ROS to the cytosol.
“Mitochondrial uncoupling and increased ROS generation activates the mitochondrial permeability transition pore (mPTP), allowing the release of ROS to the cytosol (1069).”
From Sources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation (2019) · cites this paper as [1069]
Current research has advanced knowledge on how different systems can influence NO/ROS generation from other sources, termed ROS-induced ROS release.
“Current research has greatly advanced our knowledge on how different systems can influence the NO/ROS generation from other sources in which it has been termed ROS-induced ROS release (1066, 1069).”
From Sources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation (2019) · cites this paper as [1069]
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- Authors
- Zweier, J. L., Duke, S. S., Kuppusamy, P., Sylvester, J. T., & Gabrielson, E. W.
- Affiliations
- Johns Hopkins University (US)
- Corresponding author
- Zweier, Jay L.
- Venue
- FEBS Letters
- Publisher
- Wiley
- Published
- 1989-07
- Volume
- 252
- Issue
- 1-2
- Pages
- 12-16
- ISSN
- 0014-5793 (ISSN-L)
- 1873-3468
- OpenAlex
- W2051338529
- Cited by
- 67
- References
- 18
- Citations by year
- 2024: 1 · 2018: 1 · 2016: 2 · 2015: 2 · 2014: 1 · 2013: 1 · 2012: 1
- FWCI
- 1.77
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- FEBS Letters (publishedVersion)
- PubMed (publishedVersion)
- Funders
- American Heart Association
- National Institutes of Health (HL38324)
- National Heart, Lung, and Blood Institute (HL-38324, HL-17655-13, HL41970)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Electron Spin Resonance Studies
- Health Sciences › Medicine › Biochemistry › Antioxidant Activity and Oxidative Stress
- Keywords
- hyperoxia, superoxide, hydroxyl radicals, free radical generation, pulmonary endothelial cells, superoxide dismutase, DMPO spin trapping, cell viability, oxidative cell death
- MeSH terms
- Animals, Cells, Cultured, Electron Spin Resonance Spectroscopy, Endothelium, Free Radicals, Hydroxides, Lung, Oxygen, Sheep, Superoxides
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