Superoxide production from paraquat evoked by exogenous NADPH in pulmonary endothelial cells
Abstract
Superoxide production from paraquat in a pulmonary microvascular endothelial cell (PMEC) suspension was demonstrated using 2-methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo[1,2-alpha]pyraz in-3-one (MCLA), a chemiluminescence probe, to detect superoxide anions. Increased rates of superoxide production from paraquat, which were sensitive to superoxide dismutase (SOD), required the presence of reduced nicotinamide adenine dinucleotide phosphate (NADPH) in the reaction medium, and occurred instantaneously after the addition of NADPH, which is impermeable to cell membranes. NADH as an electron donor was not as effective, and xanthine or succinate had no influence. Paraquat was anaerobically reduced in the presence of NADPH and PMECs to yield a one-electron reduced radical, and the reduction was inhibited by NADP+. Diphenyleneiodonium, an inhibitor of flavoprotein reductases, also markedly inhibited both paraquat reduction and superoxide production. These results indicate that NADPH-dependent superoxide production from paraquat probably occurs by a flavoprotein with NADPH-dependent reductase activity in cell membranes. NADPH-dependent superoxide production from paraquat was also reproduced using adherent PMECs on wells. Under these conditions, superoxide production was enhanced with agonists, including interleukin-1beta, A23187, and phorbol 12-myristate 13-acetate. The effect of the former two was blocked with staurosporine, while the latter's effect was suppressed with calyculin A.
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In herbicide paraquat poisoning, oxidative stress is a primary cause, characterized by a redox cycling mechanism involving superoxide radical anion formation triggered by exogenous NADPH in pulmonary endothelial cells.
“The primary cause of oxidative stress has also been observed in herbicide paraquat poisoning, characterized by a redox cycling mechanism involving superoxide radical anion formation triggered by exogenous NADPH in pulmonary endothelial cells (Tampo et al. 1999).”
From Reactive Oxygen Species, Toxicity, Oxidative Stress, and Antioxidants: Chronic Diseases and Aging (2023) · cites this paper as Tampo et al. 1999
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- Authors
- Tampo, Y., Tsukamoto, M., & Yonaha, M.
- Corresponding author
- Yonaha, Masanori
- Venue
- Free Radical Biology and Medicine
- Publisher
- Elsevier BV
- Published
- 1999-09
- Volume
- 27
- Issue
- 5-6
- Pages
- 588-595
- ISSN
- 0891-5849 (ISSN-L)
- OpenAlex
- W1973040816
- Cited by
- 51
- References
- 45
- Citations by year
- 2023: 2 · 2022: 1 · 2018: 2 · 2017: 2 · 2016: 1 · 2014: 2 · 2013: 1 · 2012: 1
- FWCI
- 2.10
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Free Radical Biology and Medicine (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Health Sciences › Medicine › Surgery › Paraquat toxicity studies and treatments
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Life Sciences › Immunology and Microbiology › Immunology › Neutrophil, Myeloperoxidase and Oxidative Mechanisms
- Keywords
- paraquat, superoxide production, pulmonary microvascular endothelial cells, flavoprotein reductase, diphenyleneiodonium, NADPH-dependent reductase, superoxide dismutase, chemiluminescence, IL-1β, staurosporine, calyculin A, A23187, phorbol myristate acetate, oxidative stress
- MeSH terms
- Animals, Cells, Cultured, Endothelium, Vascular, Enzyme Inhibitors, Flavoproteins, Imidazoles, Interleukin-1, Luminescent Measurements, Lung, NAD, NADP, Oxidoreductases, Paraquat, Pyrazines, Spectrophotometry, Superoxides, Superoxide Dismutase, Swine, Succinic Acid, Xanthine
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
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