Mechanisms of ascorbic acid recycling in human erythrocytes
Abstract
Vitamin C, or ascorbic acid, is efficiently recycled from its oxidized forms by human erythrocytes. In this work the dependence of this recycling on reduced glutathione (GSH) was evaluated with regard to activation of the pentose cycle and to changes in pyridine nucleotide concentrations. The two-electron-oxidized form of ascorbic acid, dehydroascorbic acid (DHA) was rapidly taken up by erythrocytes and reduced to ascorbate, which reached intracellular concentrations as high as 2 mM. In the absence of D-glucose, DHA caused dose-dependent decreases in erythrocyte GSH, NADPH, and NADH concentrations. In the presence of 5 mM D-glucose, GSH and NADH concentrations were maintained, but those of NADPH decreased. Reduction of extracellular ferricyanide by erythrocytes, which reflects intracellular ascorbate recycling, was also enhanced by D-glucose, and ferricyanide activated the pentose cycle. Diethylmaleate at concentrations up to 1 mM was found to specifically deplete erythrocyte GSH by 75-90% without causing oxidant stress in the cells. Such GSH-depleted erythrocytes showed parallel decreases in their ability to take up and reduce DHA to ascorbate, and to reduce extracellular ferricyanide. These results show that DHA reduction involves GSH-dependent activation of D-glucose metabolism in the pentose cycle, but that in the absence of D-glucose DHA reduction can also utilize NADH.
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Red blood cells have more antioxidant enzymes and send more glucose through the pentose phosphate pathway to make NADPH, which normally shields them from high-dose ascorbate.
“they have an increased level of antioxidant enzymes and enhanced glucose flux into the pentose phosphate pathway (ppp)60,61 to generate more NADPH. In a normal setting, this protects erythrocytes from high-dose ascorbate.”
From Targeting cancer vulnerabilities with high-dose vitamin C (2019) · cites this paper as [61]
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- Authors
- May, J. M., Qu, Z.-C., & Morrow, J. D.
- Affiliations
- Vanderbilt University (US)
- Corresponding author
- May, James M
- ORCID
- May, James M
- Venue
- Biochimica et Biophysica Acta (BBA) - General Subjects
- Publisher
- Elsevier BV
- Published
- 2001-10
- Volume
- 1528
- Issue
- 2-3
- Pages
- 159-166
- ISSN
- 0304-4165 (ISSN-L)
- OpenAlex
- W2084327512
- Cited by
- 81
- References
- 36
- Citations by year
- 2026: 3 · 2024: 1 · 2023: 4 · 2022: 4 · 2021: 3 · 2020: 4 · 2019: 5 · 2018: 4 · 2017: 1 · 2016: 6 · 2015: 6 · 2014: 2 · 2013: 4 · 2012: 1
- FWCI
- 4.37
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Biochimica et Biophysica Acta (BBA) - General Subjects (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health
- National Institute of Diabetes and Digestive and Kidney Diseases (R01DK050435, DK-50435)
- 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
- dehydroascorbic acid, human erythrocytes, reduced glutathione, pentose phosphate pathway, D-glucose metabolism, NADPH, NADH, ferricyanide reduction, diethyl maleate, glutathione depletion, vitamin C
- MeSH terms
- Ascorbic Acid, Dehydroascorbic Acid, Erythrocytes, Glucose, Glutathione, Humans, Maleates, NAD, NADP, Oxidation-Reduction, Pentose Phosphate Pathway, Substrate Cycling
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