Ascorbate recycling in human erythrocytes: Role of GSH in reducing dehydroascorbate
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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Within cells, DHA is quickly converted back to ascorbate while consuming GSH and NADPH.
“Inside the cell, DHA is rapidly reduced back to ascorbate at the expense of GSH and NADPH47,49.”
From Targeting cancer vulnerabilities with high-dose vitamin C (2019) · cites this paper as [49]
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Self-citationKim, Y.-S., Hwang, J.-W., Kang, S.-H., Kim, E., Jeon, Y., Jeong, J.-H., Kim, H., Moon, S.-H., Jeon, B.-T., & Park, P. (2013) · Journal of Natural Medicines · doi:10.1007/s11418-013-0786-8 · Cited by 35 · see citing works
Reduction of dehydroascorbic acid by homocysteine
Park, J. B. (2001) · Biochimica et Biophysica Acta (BBA) - General Subjects · doi:10.1016/s0304-4165(00)00186-0 · Cited by 35 · see citing works
Effect of vitamins C and E on oxidative processes in human erythrocytes
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Human erythrocytes transport dehydroascorbic acid and sugars using the same transporter complex
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Redox regulation of ascorbic acid transport: Role of transporter and intracellular sulfhydryls
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Self-citationGlut-1 explains the evolutionary advantage of the loss of endogenous vitamin C-synthesis
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Ascorbic Acid Might Play a Role in the Sclerotial Differentiation of Sclerotium rolfsii
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Recycling of the ascorbate free radical by human erythrocyte membranes
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Self-citationVitamin C recycling and function in human monocytic U-937 cells
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Antioxidant prevention of Heinz body formation and oxidative injury in cats
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Zhang, Z., Peng, L., Fu, Y., Wang, W., Wang, P., & Zhou, F. (2021) · ACS Chemical Neuroscience · doi:10.1021/acschemneuro.0c00713 · Cited by 31 · see citing works
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Paamoni-Keren, O., Silberstein, T., Burg, A., Raz, I., Mazor, M., & Saphier, O. (2007) · Archives of Gynecology and Obstetrics · doi:10.1007/s00404-006-0304-2 · Cited by 31 · see citing works
Age-Associated Oxidative Macromolecular Damages in Rat Brain Regions: Role of Glutathione Monoester
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Interplay Between Metabolic Pathways and Increased Oxidative Stress in Human Red Blood Cells
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Oxalate mediated nephronal impairment and its inhibition by c-phycocyanin: A study on urolithic rats
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Plasma Membrane Oxidoreductases: Effects on Erythrocyte Metabolism and Redox Homeostasis
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Uptake and reduction of α-lipoic acid by human erythrocytes
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The role of ascorbic acid role in the differentiation of sclerotia in Sclerotinia minor
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Crosstalk and Barriers Between the Electron Carriers of the Endoplasmic Reticulum
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Rasool, M., Ashraf, M. A. B., Malik, A., Waquar, S., Khan, S. A., Qazi, M. H., Ahmad, W., Asif, M., Khan, S. U., Zaheer, A., et al. (2017) · PLoS ONE · doi:10.1371/journal.pone.0171561 · Cited by 23 · see citing works
- 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)
- 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
- 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 › 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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