Cited by 430Nitric Oxide and Endothelin EffectsRedox biology and oxidative stressNeutrophil, Myeloperoxidase and Oxidative Mechanisms

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Transcriptional and Posttranscriptional Regulation of Endothelial Nitric Oxide Synthase Expression by Hydrogen Peroxide

Drummond, Grant R., Cai, Hua, Davis, Michael E., Ramasamy, Santhini, Harrison, David G.·2000·Circulation Research

DOI 10.1161/01.res.86.3.347

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Abstract

Abstract —Diverse stimuli, including shear stress, cyclic strain, oxidized LDL, hyperglycemia, and cell growth, modulate endothelial nitric oxide synthase (eNOS) expression. Although seemingly unrelated, these may all alter cellular redox state, suggesting that reactive oxygen intermediates might modulate eNOS expression. The present study was designed to test this hypothesis. Exposure of bovine aortic endothelial cells for 24 hours to paraquat, a superoxide (O 2 −· )–generating compound, did not affect eNOS mRNA levels. However, cotreatment with paraquat and either Cu 2+ /Zn 2+ superoxide dismutase or the superoxide dismutase mimetic tetrakis(4-benzoic acid)porphyrin chloride increased eNOS mRNA by 2.3- and 2.2-fold, respectively, implicating a role for H 2 O 2 . Direct addition of 100 and 150 μmol/L H 2 O 2 caused increases in bovine aortic endothelial cell eNOS mRNA that were dependent on concentration (ie, 3.1- and 5.2-fold increases) and time, and elevated eNOS protein expression and enzyme activity, accordingly. Nuclear run-on and 5,6-dichloro-1-β- d -ribofuranosylbenzimidazole–chase studies showed that H 2 O 2 caused a 3.0-fold increase in eNOS gene transcription and a 2.8-fold increase in eNOS mRNA half-life. Induction of eNOS by H 2 O 2 was not affected by the hydroxyl radical scavenger DMSO, mannitol, or N-tert-butyl -α-phenylnitrone, but it was inhibited by the antioxidants N -acetylcysteine, ebselen, and exogenously added catalase. Unlike H 2 O 2 , the 4.0-fold induction of eNOS by shear stress (15 dyne/cm 2 for 6 hours) was not inhibited by N -acetylcysteine or exogenous catalase. In conclusion, H 2 O 2 increases eNOS expression through transcriptional and post-transcriptional mechanisms. Although H 2 O 2 does not mediate shear-dependent eNOS regulation, it is likely to be involved in regulation of eNOS expression in response to other physiological and/or pathophysiological stimuli.

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Authors: Drummond, G. R., Cai, H., Davis, M. E., Ramasamy, S., & Harrison, D. G.
Affiliations: Emory University (US)
ORCID: Drummond, Grant R.
Cai, Hua
Davis, Michael E.
Ramasamy, Santhini
Harrison, David G.
Venue: Circulation Research
Publisher: Ovid Technologies (Wolters Kluwer Health)
Published: 2000-02-18
Volume: 86
Issue: 3
Pages: 347-354
ISSN: 0009-7330 (ISSN-L)
1524-4571
DOI: 10.1161/01.res.86.3.347
OpenAlex: W2110361956
Cited by: 430
References: 34
Citations by year: 2026: 2 · 2025: 5 · 2024: 13 · 2023: 8 · 2022: 8 · 2021: 11 · 2020: 12 · 2019: 10 · 2018: 7 · 2017: 14 · 2016: 13 · 2015: 15 · 2014: 19 · 2013: 15 · 2012: 16
FWCI: 16.31
Type: journal article
Language: en
Open access: closed
Locations: Circulation Research (publishedVersion)
PubMed (publishedVersion)
Funders: U.S. Department of Veterans Affairs
American Heart Association
National Institutes of Health (5r01hl059248-04)
National Heart, Lung, and Blood Institute (HL 58000, HL 59248, HL 39000 6, P01HL058000)
Topics: Health Sciences › Medicine › Physiology (medical) › Nitric Oxide and Endothelin Effects
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: endothelial nitric oxide synthase, hydrogen peroxide, mRNA stability, post-transcriptional regulation, shear stress, superoxide dismutase, paraquat, redox state, reactive oxygen species, antioxidant inhibition, N-acetylcysteine, ebselen, catalase, endothelial cells, oxidative stress, nitric oxide signaling
MeSH terms: Animals, Cattle, Cells, Cultured, Enzyme Induction, Humans, Hydrogen Peroxide, Paraquat, Protein Processing, Post-Translational, RNA, Messenger, Stress, Mechanical, Time Factors, Transcription, Genetic, Free Radical Scavengers, Oxidants, Nitric Oxide Synthase, RNA Stability, Nitric Oxide Synthase Type III

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Transcriptional and Posttranscriptional Regulation of Endothelial Nitric Oxide Synthase Expression by Hydrogen Peroxide

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