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Ascorbate uptake and antioxidant function in peritoneal macrophages.

James M May1, Liying Li, Zhi-Chao Qu

  • 1Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232-6303, USA. james.may@vanderbilt.edu

Archives of Biochemistry and Biophysics
|August 2, 2005
PubMed
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Activated macrophages use ascorbic acid (vitamin C) as an antioxidant to combat reactive oxygen species. Loading cells with vitamin C protects against oxidant stress and preserves alpha-tocopherol.

Area of Science:

  • Immunology
  • Cell Biology
  • Nutritional Science

Background:

  • Activated macrophages produce reactive oxygen species (ROS), which can be harmful.
  • Ascorbic acid (vitamin C) is a known antioxidant.
  • The role of ascorbic acid in macrophage antioxidant defense is not fully understood.

Purpose of the Study:

  • To investigate whether ascorbic acid functions as an antioxidant in activated macrophages.
  • To determine the mechanism of ascorbic acid uptake and its effect on oxidant stress.

Main Methods:

  • Thioglycollate-elicited murine peritoneal macrophages were cultured.
  • Ascorbate uptake and intracellular concentrations were measured.
  • Cells were activated by phagocytosis of latex particles.

Related Experiment Videos

  • Reactive oxygen species (ROS) and alpha-tocopherol levels were assessed.
  • The expression of the sodium-dependent vitamin C transporter 2 (SVCT2) was examined.

    • Main Results:

    • Macrophages contain significant intracellular ascorbate levels, regulated by a high-affinity sodium-dependent transport mechanism, likely via SVCT2.
    • Phagocytosis by activated macrophages depleted intracellular ascorbate, indicating its sensitivity to oxidant stress.
    • Ascorbate loading prevented the loss of alpha-tocopherol and the increase in ROS induced by phagocytosis.
    • Glutathione (GSH) levels were unaffected, suggesting ascorbate is more sensitive to phagocytic oxidant stress.

    Conclusions:

    • Activated macrophages utilize ascorbic acid to mitigate self-generated oxidative stress.
    • Ascorbate protects alpha-tocopherol, potentially preventing macrophage necrosis or apoptosis.
    • These findings highlight ascorbic acid's crucial role in macrophage antioxidant defense.