Hyperglycemia increases mitochondrial superoxide in retina and retinal cells
Abstract
Oxidative stress is believed to play a significant role in the development of diabetic retinopathy. In this study, we have investigated the effects of elevated glucose concentration on the production of superoxide anion by retina and retinal cells, the cellular source of the superoxide, the effect of therapies that are known to inhibit diabetic retinopathy on the superoxide production, and the role of the superoxide in cell death in elevated glucose concentration. Superoxide release was measured from retinas collected from streptozotocin-diabetic rats (2 months) treated with or without aminoguanidine, aspirin, or vitamin E, and from transformed retinal Müller cells (rMC-1) and bovine retinal endothelial cells (BREC) incubated in normal (5 mM) and high (25 mM) glucose. Diabetes (retina) or incubation in elevated glucose concentration (rMC-1 and BREC cells) significantly increased superoxide production, primarily from mitochondria, because an inhibitor of mitochondrial electron transport chain complex II normalized superoxide production. Inhibition of reduced nicotinamine adenine dinucleotide phosphate (NADPH) oxidase or nitric oxide synthase had little or no effect on the glucose-induced increase in superoxide. Treatment of diabetic animals with aminoguanidine, aspirin, or vitamin E for 2 months significantly inhibited the diabetes-induced increase in production of superoxide in the retinas. Despite the increased production of superoxide, no increase in protein carbonyls was detected in retinal proteins from animals diabetic for 2-6 months or rMC-1 cells incubated in 25 mM glucose for 5 d unless the activities of calpain or the proteosome were inhibited. Addition of copper/zinc-containing superoxide dismutase to the media of rMC-1 and BREC cells inhibited the apoptotic death caused by elevated glucose. Diabetes-like glucose concentration increases superoxide production in retinal cells, and the superoxide contributes to impaired viability and increased cell death under those circumstances. Three therapies that inhibit the development of diabetic retinopathy all inhibit superoxide production, raising a possibility that these therapies inhibit retinopathy in part by inhibiting a hyperglycemia-induced increase in superoxide production.
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In transformed retinal cells and bovine retinal endothelial cells, incubation in 25 mM vs 5 mM glucose increases superoxide production.
“In transformed retinal cells (rMC-1s) and bovine retinal endothelial cells (BRECs), incubation in 25 mM, as opposed to 5 mM, glucose increased superoxide production (87).”
From Mitochondrial Dysfunction in Diabetes: From Molecular Mechanisms to Functional Significance and Therapeutic Opportunities (2010) · cites this paper as [87]
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- Authors
- Du, Y., Miller, C. M., & Kern, T. S.
- Affiliations
- Case Western Reserve University (US)
- Corresponding author
- Kern, T.S
- ORCID
- Venue
- Free Radical Biology and Medicine
- Publisher
- Elsevier BV
- Published
- 2003-12
- Volume
- 35
- Issue
- 11
- Pages
- 1491-1499
- ISSN
- 0891-5849 (ISSN-L)
- OpenAlex
- W2043243208
- Cited by
- 355
- References
- 49
- Citations by year
- 2026: 10 · 2025: 16 · 2024: 5 · 2023: 9 · 2022: 7 · 2021: 13 · 2020: 22 · 2019: 11 · 2018: 15 · 2017: 19 · 2016: 14 · 2015: 22 · 2014: 21 · 2013: 24 · 2012: 23
- FWCI
- 11.74
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Free Radical Biology and Medicine (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Case Western Reserve University
- National Eye Institute (EY00 300, P30 EY-11373, P30EY011373)
- National Institute of Diabetes and Digestive and Kidney Diseases (P01DK057733, DK-57733)
- Topics
- Health Sciences › Medicine › Ophthalmology › Retinal Diseases and Treatments
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Clinical Biochemistry › Advanced Glycation End Products research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Keywords
- hyperglycemia, mitochondrial superoxide, diabetic retinopathy, oxidative stress, Müller cells, retinal endothelial cells, aminoguanidine, aspirin, vitamin E, superoxide dismutase, apoptosis, NADPH oxidase, nitric oxide synthase, protein carbonyls, calpain, proteasome, streptozotocin-induced diabetes
- MeSH terms
- Actins, Animals, Cattle, Cell Line, Transformed, Cell Survival, Copper, Diabetic Retinopathy, Electron Transport, Glucose, Hyperglycemia, Immunohistochemistry, Male, Mitochondria, Oxygen, Rats, Inbred Lew, Retina, Superoxides, Zinc, Cell Death, Apoptosis, Oxidative Stress, Nitric Oxide Synthase, Rats
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