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Ascorbic acid efflux and re-uptake in endothelial cells: maintenance of intracellular ascorbate

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Abstract

Entry of vitamin C or ascorbate into most tissues requires its movement across the endothelial cell barrier of vessels. If trans-cellular ascorbate movement occurs, then it should be evident as ascorbate efflux from endothelial cells. Cultured EA.926 endothelial cells that had been loaded to about 3.5 mM intracellular ascorbate lost 70–80% of ascorbate to the medium over several hours at 37°C via a non-saturable process that was insensitive to anion transport inhibitors and thiol reagents. Oxidation of this extracellular ascorbate by ascorbate oxidase or ferricyanide enhanced apparent ascorbate efflux, suggesting that efflux of the vitamin was countered in part by its re-uptake on ascorbate transporters. Although basal ascorbate efflux was not calcium-dependent, increased entry of calcium into the cells enhanced ascorbate release. These results support the hypothesis that ascorbate efflux reflects trans-endothelial cell ascorbate movement out of the blood vessel.

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Abbreviations

DHA:

Dehydroascorbic acid

DIDS:

4,4′-Diisothiocyanatostilbene-2,2′-disulfonic acid

EDTA:

Ethylenediamine-tetraacetic acid

GSH:

Reduced glutathione

Hepes:

N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid

KRH:

Krebs-Ringer Hepes

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Acknowledgments

This work was supported by NIH grant DK 50435 and by the Vanderbilt Diabetes Research and Training Center (DK 20593).

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  1. Department of Medicine, Vanderbilt University School of Medicine, 7465 Medical Research Building IV, Nashville, TN, 37232-0475, USA

    James M. May & Zhi-chao Qu

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  1. James M. May
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Correspondence to James M. May.

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May, J.M., Qu, Zc. Ascorbic acid efflux and re-uptake in endothelial cells: maintenance of intracellular ascorbate. Mol Cell Biochem 325, 79–88 (2009). https://doi.org/10.1007/s11010-008-0022-0

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