ASCORBIC ACID FUNCTION AND METABOLISM

Summary

Principal Investigator: James M May
Affiliation: Vanderbilt University
Country: USA
Abstract: Ascorbic acid, or vitamin C, is the most important water-soluble antioxidant in the human circulation. Although humans cannot synthesize ascorbate de novo, it can be efficiently recycled within cells from its oxidized product, dehydroascorbate. Our work has suggested that such ascorbate recycling is particularly important in the human erythrocyte, where such recycling can contribute both to circulating levels of the vitamin, and to protecting the erythrocyte membrane against lipid peroxidative damage. The goals of this proposal are to determine the mechanisms by which erythrocyte ascorbate recycling contributes to the defense against lipid peroxidation in blood, and to establish the extent to which this protection occurs. To achieve these goals, we will utilize techniques of fluorescence spectroscopy, EPR spectroscopy, molecular cloning, and novel biochemical assays. Whether and how ascorbate functions to protect cell membranes and lipoproteins from oxidation is relevant to mechanisms of inflammation, ageing, and atherosclerosis, especially given the fact that dietary supplementation can more than double circulating and tissue levels of the vitamin. In the first specific aim, we will address the question of how reducing equivalents from intracellular ascorbate are transferred across or into the cell membrane. In erythrocytes such transfer appears to be mediated via a transmembrane ascorbate-dependent oxidoreductase enzyme. We will first complete our studies of the kinetic mechanism of this enzyme activity, purify it from human erythrocytes, and clone its cDNA sequence from a bone marrow-derived cDNA library. We will also evaluate the role of ubiquinol-10 (coenzyme Q10) as a cofactor necessary for electron transfer by the enzyme as well as whether the enzyme is functions similarly in a cultured monocyte-derived cell. In the second specific aim-, we will determine whether and to what extent membrane-bound lipid hydroperoxides, alpha-tocopherol and ubiquinone-10 are natural acceptors of electrons derived from ascorbate within erythrocytes. In the third specific aim we will determine whether extracellular ascorbate or its lipophilic derivatives can prevent oxidant-mediated damage in erythrocyte membranes, either directly or through recycling of alpha tocopherol. We will also test the hypothesis that ascorbate-dependent recycling of alpha tocopherol in erythrocytes can replenish depleted stores of alpha tocopherol in human LDL.
Funding Period: 1995-09-30 - 2000-08-31
more information: NIH RePORT

Top Publications

  1. pmc Ascorbate reverses high glucose- and RAGE-induced leak of the endothelial permeability barrier
    M Elizabeth Meredith
    Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, TN 37232 0475, United States
    Biochem Biophys Res Commun 445:30-5. 2014
  2. pmc Role of vitamin C in the function of the vascular endothelium
    James M May
    Department of Medicine, Vanderbilt University School of Medicine, Nashville, Tennessee
    Antioxid Redox Signal 19:2068-83. 2013
  3. pmc Chelation of intracellular iron enhances endothelial barrier function: a role for vitamin C?
    James M May
    Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232 0475, United States
    Arch Biochem Biophys 500:162-8. 2010
  4. pmc Oxidized LDL up-regulates the ascorbic acid transporter SVCT2 in endothelial cells
    James M May
    Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232 0475, USA
    Mol Cell Biochem 343:217-22. 2010
  5. pmc Combined vitamin C and vitamin E deficiency worsens early atherosclerosis in apolipoprotein E-deficient mice
    Vladimir R Babaev
    Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232 0475, USA
    Arterioscler Thromb Vasc Biol 30:1751-7. 2010
  6. pmc Ascorbic acid prevents increased endothelial permeability caused by oxidized low density lipoprotein
    James M May
    Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232 0475, USA
    Free Radic Res 44:1359-68. 2010
  7. pmc Selective macrophage ascorbate deficiency suppresses early atherosclerosis
    Vladimir R Babaev
    Department of Medicine, Vanderbilt University Medical Center, Nashville, TN 37232, USA
    Free Radic Biol Med 50:27-36. 2011
  8. pmc Nitric oxide mediates tightening of the endothelial barrier by ascorbic acid
    James M May
    Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232 0475, USA
    Biochem Biophys Res Commun 404:701-5. 2011
  9. pmc Ascorbic acid prevents oxidant-induced increases in endothelial permeability
    James M May
    Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232, USA
    Biofactors 37:46-50. 2011
  10. pmc Assessing the reductive capacity of cells by measuring the recycling of ascorbic and lipoic acids
    James M May
    Department of Medicine, Vanderbilt University Medical Center, Nashville, TN, USA
    Methods Mol Biol 610:229-43. 2010

Scientific Experts

  • James M May
  • Huan Qiao
  • Vladimir R Babaev
  • Liying Li
  • M Elizabeth Meredith
  • Zhi Chao Qu
  • Sergio Fazio
  • Macrae F Linton
  • Fiona E Harrison
  • Xiumei Chi
  • Anup K Sabharwal
  • Rene Aguirre
  • Dan Su
  • Richard R Whitesell
  • Sanket Shah
  • Saul Juliao
  • Jason Bell
  • Han Asard
  • Mark J Koury

Detail Information

Publications26

  1. pmc Ascorbate reverses high glucose- and RAGE-induced leak of the endothelial permeability barrier
    M Elizabeth Meredith
    Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, TN 37232 0475, United States
    Biochem Biophys Res Commun 445:30-5. 2014
    ....
  2. pmc Role of vitamin C in the function of the vascular endothelium
    James M May
    Department of Medicine, Vanderbilt University School of Medicine, Nashville, Tennessee
    Antioxid Redox Signal 19:2068-83. 2013
    ..Although ascorbate may not be able to reverse inflammatory vascular diseases such as atherosclerosis, it may well play a role in preventing the endothelial dysfunction that is the earliest sign of many such diseases...
  3. pmc Chelation of intracellular iron enhances endothelial barrier function: a role for vitamin C?
    James M May
    Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232 0475, United States
    Arch Biochem Biophys 500:162-8. 2010
    ..These results show that chelation of intracellular iron decreases endothelial barrier permeability and implicate this mechanism in the ability of EDHB and possibly intracellular ascorbate to tighten the endothelial barrier...
  4. pmc Oxidized LDL up-regulates the ascorbic acid transporter SVCT2 in endothelial cells
    James M May
    Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232 0475, USA
    Mol Cell Biochem 343:217-22. 2010
    ..These results suggest that up-regulation of endothelial cell SVCT2 expression and function may help to maintain intracellular ascorbate during oxLDL-induced oxidative stress, and that ascorbate in turn can prevent this effect...
  5. pmc Combined vitamin C and vitamin E deficiency worsens early atherosclerosis in apolipoprotein E-deficient mice
    Vladimir R Babaev
    Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232 0475, USA
    Arterioscler Thromb Vasc Biol 30:1751-7. 2010
    ....
  6. pmc Ascorbic acid prevents increased endothelial permeability caused by oxidized low density lipoprotein
    James M May
    Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232 0475, USA
    Free Radic Res 44:1359-68. 2010
    ..These results suggest a role for endothelial cell ascorbate in ameliorating an important facet of endothelial dysfunction caused by mLDL...
  7. pmc Selective macrophage ascorbate deficiency suppresses early atherosclerosis
    Vladimir R Babaev
    Department of Medicine, Vanderbilt University Medical Center, Nashville, TN 37232, USA
    Free Radic Biol Med 50:27-36. 2011
    ....
  8. pmc Nitric oxide mediates tightening of the endothelial barrier by ascorbic acid
    James M May
    Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232 0475, USA
    Biochem Biophys Res Commun 404:701-5. 2011
    ..These results show that endothelial permeability barrier function depends on constitutive generation of NO and that ascorbate-dependent tightening of this barrier involves maintaining NO through the eNOS/guanylate cyclase pathway...
  9. pmc Ascorbic acid prevents oxidant-induced increases in endothelial permeability
    James M May
    Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232, USA
    Biofactors 37:46-50. 2011
    ..These results suggest that oxidative stress induced directly by oxidants or indirectly by glutathione depletion impairs endothelial barrier function and that intracellular ascorbate may serve to prevent this effect...
  10. pmc Assessing the reductive capacity of cells by measuring the recycling of ascorbic and lipoic acids
    James M May
    Department of Medicine, Vanderbilt University Medical Center, Nashville, TN, USA
    Methods Mol Biol 610:229-43. 2010
    ..Together, these assays provide an estimate of the ability of different cell types to recycle ascorbate and to generate intracellular reducing equivalents required to maintain the redox status of the cells...
  11. pmc Transfer of ascorbic acid across the vascular endothelium: mechanism and self-regulation
    James M May
    Department of Medicine, Vanderbilt Univ School of Medicine, Nashville, TN 37232 0475, USA
    Am J Physiol Cell Physiol 297:C169-78. 2009
    ..Since the latter effect occurred over the physiological range of ascorbate plasma concentrations, it could reflect a role for the vitamin in control of endothelial barrier function in vivo...
  12. pmc Cobalt-induced oxidant stress in cultured endothelial cells: prevention by ascorbate in relation to HIF-1alpha
    Huan Qiao
    Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232 0475, USA
    Biofactors 35:306-13. 2009
    ....
  13. pmc Oxidized lipoprotein induces the macrophage ascorbate transporter (SVCT2): protection by intracellular ascorbate against oxidant stress and apoptosis
    Xiumei Chi
    Department of Medicine, Vanderbilt University School of Medicine, 7465 Medical Research Building IV, Nashville, TN 37232 6303, USA
    Arch Biochem Biophys 485:174-82. 2009
    ..These results suggest that ascorbate protects macrophages against oxLDL-induced oxidant stress and subsequent apoptotic death without impairing their function...
  14. pmc Macrophage differentiation increases expression of the ascorbate transporter (SVCT2)
    Huan Qiao
    Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232 6303, USA
    Free Radic Biol Med 46:1221-32. 2009
    ....
  15. ncbi Ascorbic acid decreases oxidant stress in endothelial cells caused by the nitroxide tempol
    James M May
    Department of Medicine, 754 Preston Research Building, Vanderbilt University School of Medicine, Nashville TN 37232 6303, USA
    Free Radic Res 39:195-202. 2005
    ..These results show that whereas Tempol may scavenge other more toxic radicals, care must be taken to ensure that it does not itself induce an oxidant stress, especially with regard to depletion of ascorbic acid...
  16. ncbi Ascorbate uptake and antioxidant function in peritoneal macrophages
    James M May
    Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232 6303, USA
    Arch Biochem Biophys 440:165-72. 2005
    ..These results suggest that activated macrophages can use ascorbate to lessen self-generated oxidant stress and spare alpha-tocopherol, which may protect these long-lived cells from necrosis or apoptosis...
  17. ncbi Macrophage uptake and recycling of ascorbic acid: response to activation by lipopolysaccharide
    James M May
    Department of Medicine, 715 Preston Research Building, Vanderbilt University School of Medicine, Nashville, TN 37232 6303, USA
    Free Radic Biol Med 39:1449-59. 2005
    ....
  18. ncbi Human erythrocyte membranes contain a cytochrome b561 that may be involved in extracellular ascorbate recycling
    Dan Su
    Department of Biochemistry, University of Nebraska, Lincoln, Nebraska 68588, USA
    J Biol Chem 281:39852-9. 2006
    ..These results suggest that the DCytb present in human erythrocytes may contribute to their ability to reduce extracellular monodehydroascorbate...
  19. pmc Maturational loss of the vitamin C transporter in erythrocytes
    James M May
    Department of Medicine, Vanderbilt University School of Medicine, 715 Preston Research Building, Nashville, TN 37232 6303, USA
    Biochem Biophys Res Commun 360:295-8. 2007
    ..Rates of specific ascorbate transport correlated with the presence of the SVCT2. These results show that mature erythrocytes fail to concentrate ascorbate due to the loss of SVCT2 during maturation in the bone marrow...
  20. ncbi alpha-Lipoic acid and ascorbate prevent LDL oxidation and oxidant stress in endothelial cells
    Anup K Sabharwal
    Department of Medicine, Vanderbilt University School of Medicine, 7465 Medical Research Building IV, Nashville, TN 37232 0475, USA
    Mol Cell Biochem 309:125-32. 2008
    ..Both LA and ascorbate protect endothelial cells and LDL from either intra- or extracellular oxidant stress, but that LA does not spare ascorbate in oxidatively stressed cells...
  21. pmc Development of ascorbate transporters in brain cortical capillary endothelial cells in culture
    Huan Qiao
    Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232 0475, USA
    Brain Res 1208:79-86. 2008
    ....
  22. pmc Ascorbic acid uptake and regulation of type I collagen synthesis in cultured vascular smooth muscle cells
    Huan Qiao
    Department of Medicine, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, USA
    J Vasc Res 46:15-24. 2009
    ..Since inflammation is associated with oxidant stress, we examined the uptake and cellular effects of the antioxidant vitamin ascorbic acid in cultured A10 vascular smooth muscle cells...
  23. pmc Inflammation in the vascular bed: importance of vitamin C
    Rene Aguirre
    Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232 6303, USA
    Pharmacol Ther 119:96-103. 2008
    ..Although further studies of ascorbate function in these cell types and in novel animal models are needed, available evidence generally supports a salutary role for this vitamin in ameliorating the earliest stages of atherosclerosis...
  24. pmc Ascorbic acid efflux and re-uptake in endothelial cells: maintenance of intracellular ascorbate
    James M May
    Department of Medicine, Vanderbilt University School of Medicine, 7465 Medical Research Building IV, Nashville, TN 37232 0475, USA
    Mol Cell Biochem 325:79-88. 2009
    ..These results support the hypothesis that ascorbate efflux reflects trans-endothelial cell ascorbate movement out of the blood vessel...
  25. pmc Vitamin C function in the brain: vital role of the ascorbate transporter SVCT2
    Fiona E Harrison
    Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232 6303, USA
    Free Radic Biol Med 46:719-30. 2009
    ....
  26. ncbi Transport and intracellular accumulation of vitamin C in endothelial cells: relevance to collagen synthesis
    James M May
    Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232 6303, USA
    Arch Biochem Biophys 434:178-86. 2005
    ..Since such concentrations can only be generated by the ascorbate transporter, these results show the necessity of transport for this crucial function of the vitamin in endothelium...