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Studies with low micromolar levels of ascorbic and dehydroascorbic acid fail to unravel a preferential route for vitamin C uptake and accumulation in U937 cells

  • Catia Azzolini (a1), Mara Fiorani (a1), Andrea Guidarelli (a1) and Orazio Cantoni (a1)
Abstract

Mammalian cells accumulate vitamin C either as ascorbic acid (AA), via Na+–AA co-transport, or dehydroascorbic acid (DHA, the oxidation product of AA), via facilitative hexose transport. As the latter, unlike the former, is a high-capacity transport mechanism, cultured cells normally accumulate greater levels of vitamin C when exposed to increasing concentrations of DHA as compared with AA. We report herein similar results using the U937 cell clone used in our laboratory only under conditions in which DHA and AA are used at concentrations greater than 50–60 μm. Below 60 μm, i.e. at levels in which AA is normally found in most biological fluids, AA and DHA are in fact taken up with identical rates and kinetics. Consequently, extracellular oxidation of AA switches the mode of uptake with hardly any effect on the net amount of vitamin C accumulated. As a final note, under these conditions, neither AA nor DHA causes detectable toxicity or any change in the redox status of the cells, as assessed by the reduced glutathione/reduced pyridine nucleotide pool. These findings therefore imply that some cell types do not have a preferential route for vitamin C accumulation, and that the uptake mechanism is uniquely dependent on the extracellular availability of AA v. DHA.

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      Studies with low micromolar levels of ascorbic and dehydroascorbic acid fail to unravel a preferential route for vitamin C uptake and accumulation in U937 cells
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      Studies with low micromolar levels of ascorbic and dehydroascorbic acid fail to unravel a preferential route for vitamin C uptake and accumulation in U937 cells
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Corresponding author
*Corresponding author: Dr M. Fiorani, fax +39 722 305324, email mara.fiorani@uniurb.it
References
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1 Rose, RC (1987) Solubility properties of reduced and oxidized ascorbate as determinants of membrane permeation. Biochim Biophys Acta 924, 254256.
2 Daruwala, R, Song, J, Koh, WS, et al. (1999) Cloning and functional characterization of the human sodium-dependent vitamin C transporters hSVCT1 and hSVCT2. FEBS Lett 460, 480484.
3 Liang, WJ, Johnson, D & Jarvis, SM (2001) Vitamin C transport systems of mammalian cells. Mol Membr Biol 18, 8795.
4 Tsukaguchi, H, Tokui, T, Mackenzie, B, et al. (1999) A family of mammalian Na+-dependent l-ascorbic acid transporters. Nature 399, 7075.
5 Wang, H, Dutta, B, Huang, W, et al. (1999) Human Na(+)-dependent vitamin C transporter 1 (hSVCT1): primary structure, functional characteristics and evidence for a non-functional splice variant. Biochim Biophys Acta 1461, 19.
6 Dhariwal, KR, Harizell, WO & Levine, M (1991) Ascorbic acid and dehydroascorbic acid measurements in human plasma and serum. Am J Clin Nutr 54, 712716.
7 Canoy, D, Wareham, N, Welch, A, et al. (2005) Plasma ascorbic acid concentrations and fat distribution in 19 068 British men and women in the European Prospective Investigation into Cancer and Nutrition Norfolk cohort study. Am J Clin Nutr 82, 12031209.
8 Corti, A, Casini, AF & Pompella, A (2010) Cellular pathways for transport and efflux of ascorbate and dehydroascorbate. Arch Biochem Biophys 500, 107115.
9 Wilson, JX (2005) Regulation of vitamin C transport. Annu Rev Nutr 25, 105125.
10 Lane, DJ & Lawen, A (2009) Ascorbate and plasma membrane electron transport–enzymes vs efflux. Free Radic Biol Med 47, 485495.
11 Vera, JC, Rivas, CI, Zhang, RH, et al. (1994) Human HL-60 myeloid leukemia cells transport dehydroascorbic acid via the glucose transporters and accumulate reduced ascorbic acid. Blood 84, 16281634.
12 Guaiquil, VH, Farber, CM, Golde, DW, et al. (1997) Efficient transport and accumulation of vitamin C in HL-60 cells depleted of glutathione. J Biol Chem 272, 99159921.
13 Nualart, FJ, Rivas, CI, Montecinos, VP, et al. (2003) Recycling of vitamin C by a bystander effect. J Biol Chem 278, 1012810133.
14 Astuya, A, Caprile, T, Castro, M, et al. (2005) Vitamin C uptake and recycling among normal and tumor cells from the central nervous system. J Neurosci Res 79, 146156.
15 Linster, CL & Van Schaftingen, E (2007) Vitamin C. Biosynthesis, recycling and degradation in mammals. FEBS J 274, 122.
16 Sagun, KC, Cárcamo, JM & Golde, DW (2005) Vitamin C enters mitochondria via facilitative glucose transporter 1 (Glut1) and confers mitochondrial protection against oxidative injury. FASEB J 19, 16571667.
17 Rumsey, SC, Kwon, O, Xu, GW, et al. (1997) Glucose transporter isoforms GLUT1 and GLUT3 transport dehydroascorbic acid. J Biol Chem 272, 1898218989.
18 Daskalopoulos, R, Korcok, J, Tao, L, et al. (2002) Accumulation of intracellular ascorbate from dehydroascorbic acid by astrocytes is decreased after oxidative stress and restored by propofol. Glia 39, 124132.
19 Montel-Hagen, A, Kinet, S, Manel, N, et al. (2008) Erythrocyte Glut1 triggers dehydroascorbic acid uptake in mammals unable to synthesize vitamin C. Cell 132, 10391048.
20 Montel-Hagen, A, Sitbon, M & Taylor, N (2009) Erythroid glucose transporters. Curr Opin Hematol 16, 165172.
21 Malo, C & Wilson, JX (2000) Glucose modulates vitamin C transport in adult human small intestinal brush border membrane vesicles. J Nutr 130, 6369.
22 Malo, C & Wilson, JX (2001) Transport of ascorbic acid and dehydroascorbic acid in rat kidney cortex. FASEB J 15, A838.
23 Chen, Q, Espey, MG, Sun, AY, et al. (2007) Ascorbate in pharmacologic concentrations selectively generates ascorbate radical and hydrogen peroxide in extracellular fluid in vivo. Proc Natl Acad Sci U S A 104, 87498754.
24 Sestili, P, Brandi, G, Brambilla, L, et al. (1996) Hydrogen peroxide mediates the killing of U937 tumor cells elicited by pharmacologically attainable concentrations of ascorbic acid: cell death prevention by extracellular catalase or catalase from cocultured erythrocytes or fibroblasts. J Pharmacol Exp Ther 277, 17191725.
25 Savini, I, Duflot, S & Avigliano, L (2000) Dehydroascorbic acid uptake in a human keratinocyte cell line (HaCaT) is glutathione-independent. Biochem J 345, 665672.
26 Perez-Cruz, I, Carcamo, JM & Golde, DW (2003) Vitamin C inhibits FAS-induced apoptosis in monocytes and U937 cells. Blood 102, 336343.
27 Beutler, E (1984) Red Cell Metabolism: A Manual of Biochemical Methods, 3rd ed., pp. 131134. New York, NY: Grüne & Stratton.
28 Stocchi, V, Cucchiarini, L, Magnani, M, et al. (1985) Simultaneous extraction and reverse-phase high-performance liquid chromatographic determination of adenine and pyridine nucleotides in human red blood cells. Anal Biochem 146, 118124.
29 May, JM, Mendiratta, S, Qu, ZC, et al. (1999) Vitamin C recycling and function in human monocytic U-937 cells. Free Radic Biol Med 26, 15131523.
30 Park, JB & Levine, M (2000) Intracellular accumulation of ascorbic acid is inhibited by flavonoids via blocking of dehydroascorbic acid and ascorbic acid uptakes in HL-60, U937 and Jurkat cells. J Nutr 130, 12971302.
31 Laggner, H & Goldenberg, H (2000) Interaction of respiratory burst and uptake of dehydroascorbic acid in differentiated HL-60 cells. Biochem J 345, 195200.
32 Bánhegyi, G, Marcolongo, P, Puskás, F, et al. (1998) Dehydroascorbate and ascorbate transport in rat liver microsomal vesicles. J Biol Chem 273, 27582762.
33 Frei, B, England, L & Ames, BN (1989) Ascorbate is an outstanding antioxidant in human blood plasma. Proc Natl Acad Sci U S A 86, 63776381.
34 Beyer, RE (1994) The role of ascorbate in antioxidant protection of biomembranes: interaction with vitamin E and coenzyme Q. J Bioenerg Biomembr 26, 349358.
35 Meister, A (1994) Glutathione-ascorbic acid antioxidant system in animals. J Biol Chem 269, 93979400.
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British Journal of Nutrition
  • ISSN: 0007-1145
  • EISSN: 1475-2662
  • URL: /core/journals/british-journal-of-nutrition
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