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A Rapid and Specific Microplate Assay for the Determination of Intra- and Extracellular Ascorbate in Cultured Cells
11:56

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Published on: April 11, 2014

Ascorbic acid prevents oxidant-induced increases in endothelial permeability.

James M May1, Zhi-Chao Qu

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

Biofactors (Oxford, England)
|February 18, 2011
PubMed
Summary

Oxidative stress damages the endothelial barrier, increasing permeability. Ascorbic acid (vitamin C) can prevent this damage by maintaining barrier integrity, protecting against oxidative stress.

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Area of Science:

  • Cell biology
  • Biochemistry
  • Physiology

Background:

  • Oxidative stress acutely increases vascular endothelial permeability to large molecules.
  • Ascorbic acid (vitamin C) is an antioxidant known to tighten the endothelial permeability barrier.

Purpose of the Study:

  • To investigate whether ascorbic acid can prevent the increase in endothelial permeability caused by cellular oxidative stress.
  • To determine the role of intracellular ascorbate in maintaining endothelial barrier function under oxidative stress.

Main Methods:

  • EA.hy926 endothelial cells cultured on filter inserts were treated with hydrogen peroxide (H2O2), menadione, or buthionine sulfoximine to induce oxidative stress.
  • Endothelial permeability was assessed using radiolabeled inulin.
  • Cells were pre-treated with dehydroascorbate for short-term ascorbate loading, and nonphysiologic antioxidants (dithiothreitol, tempol) were also tested.

Main Results:

  • Induction of oxidative stress significantly increased endothelial permeability to radiolabeled inulin.
  • Short-term ascorbate loading prevented the increase in endothelial permeability induced by the oxidative agents.
  • Nonphysiologic antioxidants also demonstrated a protective effect against increased endothelial barrier permeability.

Conclusions:

  • Oxidative stress, whether induced directly by oxidants or indirectly by glutathione depletion, impairs endothelial barrier function.
  • Intracellular ascorbate plays a crucial role in preventing oxidative stress-induced damage to the endothelial barrier.
  • Ascorbic acid represents a potential therapeutic agent for conditions involving compromised endothelial barrier function due to oxidative stress.