Cited by 200Vitamin C and Antioxidants ResearchRedox biology and oxidative stressAntioxidant Activity and Oxidative Stress

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Ascorbate recycling in human erythrocytes: Role of GSH in reducing dehydroascorbate

May, James M., Qu, Zhi-Chao, Whitesell, Richard R., Cobb, Charles E.·1996·Free Radical Biology and Medicine

DOI 10.1016/0891-5849(95)02130-2

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Abstract

Human erythrocytes regenerate ascorbate from its oxidized product, dehydroascorbate. The extent to which such ascorbate recycling occurs by a GSH-dependent mechanism was investigated. In the presence of glucose, erythrocytes took up over 90% of extracellular [14C]dehydroascorbate and rapidly converted it to [14C]ascorbate, which was trapped within the cells. Dehydroascorbate uptake and reduction was not associated with generation of a monoascorbyl free radical intermediate. Uptake and reduction of dehydroascorbate by glucose-depleted erythrocytes coordinately decreased GSH and raised GSSG concentrations in erythrocytes. This effect was reversed by D-glucose, but not by L-lactate. Conversely, depletion of cellular GSH decreased the ability of cells to recycle dehydroascorbate to ascorbate, as reflected in the extent to which cells were able to reduce extracellular ferricyanide. Monoascorbyl free radical was formed during the reduction of extracellular ferricyanide, indicating that one electron transfer steps were involved in this process. In GSH-depleted cells, addition of L-lactate as an energy source for glycolysis-dependent NADH regeneration did cause a partial recovery of the ability of cells to reduce ferricyanide. However, in resealed erythrocyte ghosts containing either 4 mM GSH or 400 mu M NADH, only the GSH-containing ghosts supported regeneration of ascorbate from added dehydroascorbate. These results suggest that in human erythrocytes ascorbate regeneration from dehydroascorbate is largely GSH dependent, and that it occurs through either enzymatic or nonenzymatic reactions not involving the monoascorbyl free radical.

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Authors: May, J. M., Qu, Z.-C., Whitesell, R. R., & Cobb, C. E.
Affiliations: Vanderbilt University (US)
Corresponding author: May, James M.
ORCID: May, James M.
Venue: Free Radical Biology and Medicine
Publisher: Elsevier BV
Published: 1996
Volume: 20
Issue: 4
Pages: 543-551
ISSN: 0891-5849 (ISSN-L)
DOI: 10.1016/0891-5849(95)02130-2
OpenAlex: W2025497222
Cited by: 200
References: 27
Citations by year: 2026: 1 · 2025: 4 · 2024: 5 · 2023: 2 · 2022: 1 · 2021: 7 · 2020: 4 · 2019: 4 · 2018: 2 · 2017: 3 · 2016: 6 · 2015: 4 · 2014: 5 · 2013: 8 · 2012: 5
FWCI: 10.16
Type: journal article
Language: en
Open access: closed
Licence: www.elsevier.com/tdm/userlicense/1.0/ (tdm)
www.elsevier.com/legal/tdmrep-license (tdm)
Locations: Free Radical Biology and Medicine (publishedVersion)
PubMed (publishedVersion)
Funders: National Institutes of Health (1s10rr004075-01)
National Heart, Lung, and Blood Institute (HL34737)
National Institute of Diabetes and Digestive and Kidney Diseases (DK-26657, P30DK026657)
National Center for Research Resources (RR 04075)
Topics: Health Sciences › Nursing › Nutrition and Dietetics › Vitamin C and Antioxidants Research
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
Health Sciences › Medicine › Biochemistry (medical) › Antioxidant Activity and Oxidative Stress
Keywords: ascorbate recycling, dehydroascorbic acid, glutathione, human erythrocytes, ascorbate regeneration, ferricyanide reduction, NADH regeneration, glucose, L-lactate, glutathione disulfide, erythrocyte ghosts, single electron transfer
MeSH terms: Ascorbic Acid, Dehydroascorbic Acid, Erythrocytes, Ferricyanides, Glutathione, Humans, Oxidation-Reduction

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Ascorbate recycling in human erythrocytes: Role of GSH in reducing dehydroascorbate

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