Cited by 316Antioxidant Activity and Oxidative StressVitamin C and Antioxidants ResearchRedox biology and oxidative stress

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Protection and Recycling of α-Tocopherol in Human Erythrocytes by Intracellular Ascorbic Acid

May, James M., Qu, Zhi-chao, Mendiratta, Shalu·1998·Archives of Biochemistry and Biophysics

DOI 10.1006/abbi.1997.0473

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Abstract

Ascorbic acid can recycle alpha-tocopherol from the tocopheroxyl free radical in lipid bilayers and in micelles, but such recycling has not been demonstrated to occur across cell membranes. In this work the ability of intracellular ascorbate to protect and to recycle alpha-tocopherol in intact human erythrocytes and erythrocyte ghosts was investigated. In erythrocytes that were 80% depleted of intracellular ascorbate by treatment with the nitroxide Tempol, both 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) and ferricyanide oxidized alpha-tocopherol to a greater extent than in cells not depleted of ascorbate. In contrast, in erythrocytes in which the intracellular ascorbate concentration had been increased by loading with dehydroascorbate, loss of alpha-tocopherol was less with both oxidants than in control cells. Protection against AAPH-induced oxidation of alpha-tocopherol was not prevented by extracellular ascorbate oxidase, indicating that the protection was due to intracellular and not to extracellular ascorbate. Incubation of erythrocytes with lecithin liposomes also generated an oxidant stress, which caused lipid peroxidation in the liposomes and depleted erythrocyte alpha-tocopherol, leading to hemolysis. Ascorbate loading of the erythrocytes delayed liposome oxidation and decreased loss of alpha-tocopherol from both cells and from alpha-tocopherol-loaded liposomes. When erythrocyte ghosts were resealed to contain ascorbate and challenged with free radicals generated by AAPH outside the ghosts, intravesicular ascorbate was totally depleted over 1 h of incubation, whereas alpha-tocopherol decreased only after ascorbate was substantially oxidized. These results suggest that ascorbate within the erythrocyte protects alpha-tocopherol in the cell membrane by a direct recycling mechanism.

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Authors: May, J. M., Qu, Z.-C., & Mendiratta, S.
Affiliations: Vanderbilt University (US)
Corresponding author: May, James M.
ORCID: May, James M.
Venue: Archives of Biochemistry and Biophysics
Publisher: Elsevier BV
Published: 1998-01
Volume: 349
Issue: 2
Pages: 281-289
ISSN: 0003-9861 (ISSN-L)
DOI: 10.1006/abbi.1997.0473
OpenAlex: W1999371223
Cited by: 316
References: 49
Citations by year: 2026: 1 · 2025: 5 · 2024: 3 · 2023: 8 · 2022: 8 · 2021: 3 · 2020: 9 · 2019: 3 · 2018: 6 · 2017: 6 · 2016: 6 · 2015: 12 · 2014: 12 · 2013: 13 · 2012: 14
FWCI: 11.18
Type: journal article
Language: en
Open access: closed
Licence: www.elsevier.com/tdm/userlicense/1.0/ (tdm)
Locations: Archives of Biochemistry and Biophysics (publishedVersion)
PubMed (publishedVersion)
Funders: National Institute of Diabetes and Digestive and Kidney Diseases (R01DK050435, DK-50435)
Topics: Health Sciences › Medicine › Biochemistry (medical) › Antioxidant Activity and Oxidative Stress
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
Keywords: alpha-tocopherol, vitamin C, tocopheroxyl radical, human erythrocytes, erythrocyte ghosts, lipid peroxidation, ascorbate depletion, dehydroascorbic acid, ferricyanide, lecithin liposomes, hemolysis, plasma membrane
MeSH terms: Amidines, Antioxidants, Ascorbic Acid, Cyclic N-Oxides, Dehydroascorbic Acid, Erythrocyte Membrane, Erythrocytes, Ferricyanides, Glutathione, Humans, Kinetics, Liposomes, Oxidation-Reduction, Phosphatidylcholines, Spin Labels, Vitamin E, Oxidants

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Protection and Recycling of α-Tocopherol in Human Erythrocytes by Intracellular Ascorbic Acid

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