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Related Experiment Video

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A Rapid and Specific Microplate Assay for the Determination of Intra- and Extracellular Ascorbate in Cultured Cells
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Ascorbic acid attenuates endothelial permeability triggered by cell-free hemoglobin.

Jamie L Kuck1, Julie A Bastarache2, Ciara M Shaver1

  • 1Division of Allergy, Pulmonary and Critical Care Medicine, Department of Medicine, Vanderbilt University Medical Center, Nashville, TN, USA.

Biochemical and Biophysical Research Communications
|November 14, 2017
PubMed
Summary
This summary is machine-generated.

Cell-free hemoglobin (CFH) in sepsis increases endothelial permeability by depleting Vitamin C (ascorbate). Supplementing ascorbate can protect against this damage, offering a potential therapeutic strategy for sepsis-induced organ dysfunction.

Keywords:
Ascorbic acidCell-free hemoglobinEndothelial permeabilityOxidative stress

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Area of Science:

  • Endothelial biology
  • Sepsis pathophysiology
  • Oxidative stress

Background:

  • Endothelial permeability is crucial in sepsis, but treatments targeting known mediators have failed.
  • Cell-free hemoglobin (CFH) is elevated in sepsis, linked to organ dysfunction and oxidant injury.
  • Vitamin C (ascorbate) may offer endothelial protection in sepsis.

Purpose of the Study:

  • To investigate if high cell-free hemoglobin (CFH) levels in sepsis disrupt endothelial barrier integrity.

Main Methods:

  • Human umbilical vein endothelial cells (HUVECs) were treated with CFH +/- ascorbate.
  • Monolayer permeability assessed via ECIS and 14C-inulin transfer.
  • Cell viability and intracellular ascorbate levels were measured.

Main Results:

  • CFH dose-dependently increased endothelial permeability (50% with 1 mg/ml CFH) without affecting cell viability.
  • CFH significantly reduced intracellular ascorbate levels.
  • Ascorbate pre-treatment attenuated CFH-induced permeability.

Conclusions:

  • Cell-free hemoglobin (CFH) increases endothelial permeability partly by depleting intracellular ascorbate.
  • Ascorbate supplementation may mitigate CFH-induced permeability increases.
  • This suggests a potential therapeutic role for ascorbate in sepsis.