Oxidative Stress and Increased eNOS and NADPH Oxidase Expression in Mouse Microvessel Endothelial Cells

Ding, Hong; Aljofan, Mohamad; Triggle, Chris R.

doi:10.1002/jcp.21063

Cited by 107Nitric Oxide and Endothelin EffectsRedox biology and oxidative stressNeutrophil, Myeloperoxidase and Oxidative Mechanisms

Oxidative Stress and Increased eNOS and NADPH Oxidase Expression in Mouse Microvessel Endothelial Cells

Ding, Hong, Aljofan, Mohamad, Triggle, Chris R.·2007·Journal of Cellular Physiology

DOI 10.1002/jcp.21063

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Abstract

Elevated oxidative stress plays a key role in diabetes-associated vascular disease. In this study, we tested the hypothesis that high glucose-induced oxidative stress was associated with changes in the expression of NADPH oxidase, superoxide dismutase (SOD) and endothelial nitric oxide synthase (eNOS). Oxidative stress was assessed in cell cultures of mouse microvessel endothelial cells (MMECs) by fluorescence labelling with dihydroethidium, lucigenin-enhanced chemiluminescence and determining NADPH oxidase subunit and eNOS expression with real-time polymerase chain reaction protocol and Western blotting. Oxidative stress and expression of the NADPH oxidase subunit, p22phox, were both increased, SOD1 and 3 expression lowered and eNOS significantly elevated in MMECs treated with 40 mM glucose for 72 h compared to low glucose medium. Oxidative stress, p22phox mRNA, eNOS mRNA, and protein were lowered by concurrent incubation with sepiapterin. When eNOS protein expression in endothelial cells was significantly decreased by eNOS siRNA treatment, superoxide generation was significantly higher in the MMECs grown in low glucose, but reduced in those grown in high glucose for 72 h. Thus, exposure of MMECs to high glucose results in increased oxidative stress that is associated with increased eNOS and NADPH oxidase subunit expression, notably p22phox, and decreased expression of SOD1 and 3.

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Metformin and other drugs used for T2DM, like SGLT2 inhibitors, may also reduce hyperglycemia-induced elevated endothelial ROS, independent of negative effects on complex 1, such as via inhibition of NADPH oxidase or blocking entry of glucose into the endothelium.
“Metformin and other drugs used for T2DM, like the SGLT2 inhibitors, may also reduce hyperglycemia-induced elevated endothelial ROS, independent of negative effects on complex 1, such as via inhibition of NADPH oxidase or blocking the entry of glucose into the endothelium (32, 107–109, 187, 188)”
From A Critical Review of the Evidence That Metformin Is a Putative Anti-Aging Drug That Enhances Healthspan and Extends Lifespan (2021) · cites this paper as [108]

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Authors: Ding, H., Aljofan, M., & Triggle, C. R.
Affiliations: RMIT University (AU)
Corresponding author: Ding, Hong
ORCID: Ding, Hong
Aljofan, Mohamad
Triggle, Chris R.
Venue: Journal of Cellular Physiology
Publisher: Wiley
Published: 2007-04-18
Volume: 212
Issue: 3
Pages: 682-689
ISSN: 0021-9541 (ISSN-L)
1097-4652
DOI: 10.1002/jcp.21063
OpenAlex: W2041427283
Cited by: 107
References: 41
Citations by year: 2026: 1 · 2024: 2 · 2023: 5 · 2022: 2 · 2021: 8 · 2020: 7 · 2019: 10 · 2018: 8 · 2017: 7 · 2016: 2 · 2015: 5 · 2014: 9 · 2013: 8 · 2012: 10
FWCI: 3.12
Type: journal article
Language: en
Open access: closed
Licence: onlinelibrary.wiley.com/termsAndConditions#vor (vor)
Locations: Journal of Cellular Physiology (publishedVersion)
PubMed (publishedVersion)
RMIT Research Repository (RMIT University Library) (submittedVersion)
Figshare (submittedVersion)
Funders: National Institutes of Health
Topics: Health Sciences › Medicine › Physiology › 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: oxidative stress, endothelial nitric oxide synthase, NADPH oxidase, p22phox, superoxide dismutase, high glucose, microvascular endothelial cells, sepiapterin, superoxide production
MeSH terms: Superoxide Dismutase-1, NADPH Oxidase 4, Acetophenones, Superoxide Dismutase 2, Alkaloids, Animals, Cells, Cultured, Cytochrome b Group, Dose-Response Relationship, Drug, Enzyme Inhibitors, Glucose, Microcirculation, Nitric Oxide, Protein Kinase C, Pterins, RNA, Messenger, Superoxides, Superoxide Dismutase, Transfection, Gene Expression Regulation, Enzymologic, Oxidative Stress, NADPH Oxidases, NG-Nitroarginine Methyl Ester, RNA Interference, RNA, Small Interfering, Endothelial Cells, Mice, Nitric Oxide Synthase Type II, Nitric Oxide Synthase Type III, Benzophenanthridines

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Oxidative Stress and Increased eNOS and NADPH Oxidase Expression in Mouse Microvessel Endothelial Cells

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