title: Efflux of hepatic ascorbate: a potential contributor to the maintenance of plasma vitamin C.
abstract: Ascorbate (AH, the reduced form of vitamin C) is an important radical scavenger and antioxidant in human plasma; the resulting ascorbyl radical can disproportionate to AH and dehydroascorbic acid (DHA). Here we address potential maintenance mechanism(s) for extracellular AH by examining the ability of cells to convert extracellularly presented DHA to AH. DHA was rapidly transported into human liver (HepG2), endothelial and whole blood cells in vitro by plasma membrane glucose transporters and reduced intracellularly. Liver cells displayed the highest capacity to release the intracellularly accumulated AH. The proteins responsible for DHA uptake and AH release could be distinguished by inhibitor studies. Thus, unlike DHA uptake, AH efflux was largely insensitive to cytochalasin B and thiol-reactive agents but was inhibited by phloretin, 4,4'-di-isothiocyanostilbene-2,2'-disulphonate and isoascorbate. Efflux of AH from cells was temperature-sensitive and saturable with a low affinity (millimolar, intracellular) for AH. In addition to isolated liver cells, perfusion of intact rat and guinea-pig liver with DHA resulted in AH in the circulating perfusate. Our results show that hepatocytes take up and reduce DHA and subsequently release part of the AH formed, probably via a membrane transporter. By converting extracellular DHA to extracellular AH, the liver might contribute to the maintenance of plasma AH, a process that could be important under conditions of oxidative stress.
authors.affiliation: Biochemistry Group, The Heart Research Institute, 145 Missenden Road, Camperdown, Sydney, NSW 2050, Australia. biochemistry@hri.org.au
authors.forename: J M
authors.initials: JM
authors.lastname: Upston
authors.forename: A
authors.initials: A
authors.lastname: Karjalainen
authors.forename: F L
authors.initials: FL
authors.lastname: Bygrave
authors.forename: R
authors.initials: R
authors.lastname: Stocker
mesh_terms: D000818:Animals; D001205:Ascorbic Acid / Q000097:blood* / Q000378:metabolism*; D001692:Biological Transport; D001773:Blood Cells / Q000378:metabolism; D002460:Cell Line; D003683:Dehydroascorbic Acid / Q000008:administration & dosage / Q000378:metabolism*; D004579:Electron Transport; D006168:Guinea Pigs; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D008297:Male; D008954:Models, Biological; D009004:Monosaccharide Transport Proteins / Q000378:metabolism; D010084:Oxidation-Reduction; D051381:Rats; D017208:Rats, Wistar; D013696:Temperature
chemical_list: D009004:Monosaccharide Transport Proteins; D001205:Ascorbic Acid; D003683:Dehydroascorbic Acid