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Peroxynitrite causes endothelial cell monolayer barrier dysfunction.

J L Knepler1, L N Taher, M P Gupta

  • 1Department of Medicine, Richard L. Roudebush Veterans Affairs and Indiana University Medical Centers, Indianapolis, Indiana 46202, USA.

American Journal of Physiology. Cell Physiology
|August 15, 2001
PubMed
Summary
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Peroxynitrite (ONOO(-)) directly damages endothelial cells, causing barrier dysfunction by altering cytoskeletal proteins like actin and beta-catenin. This finding is crucial for understanding lung injury mechanisms.

Area of Science:

  • Endothelial cell biology
  • Oxidative stress research
  • Pulmonary vascular physiology

Background:

  • Nitric oxide (NO) normally protects pulmonary artery endothelial cells (PAEC) from hydrogen peroxide (H2O2)-induced barrier dysfunction.
  • NO can react with superoxide (O2-) to form peroxynitrite (ONOO-), a potent oxidant.

Purpose of the Study:

  • To investigate the hypothesis that peroxynitrite (ONOO-) causes PAEC monolayer barrier dysfunction.
  • To determine the effects of ONOO- on endothelial cell cytoskeletal proteins and overall barrier integrity.

Main Methods:

  • PAEC were treated with ONOO- or SIN-1 (a ONOO- generator).
  • Albumin clearance was measured to assess barrier function.
  • Immunofluorescent confocal imaging was used to examine cytoskeletal protein (actin, beta-catenin) architecture.

Related Experiment Videos

  • Tyrosine nitration and protein oxidation were analyzed.

    • Main Results:

    • Both ONOO- and SIN-1 increased albumin clearance, indicating barrier dysfunction, without causing cytotoxicity.
    • ONOO- and SIN-1 altered the actin and beta-catenin cytoskeletal architecture.
    • ONOO- induced barrier dysfunction was partially reversible and attenuated by cysteine.
    • Tyrosine residues on beta-catenin and actin were nitrated, and proteins were oxidized.

    Conclusions:

    • Peroxynitrite (ONOO-) directly induces PAEC monolayer barrier dysfunction.
    • ONOO- mediated barrier dysfunction involves alterations to endothelial cytoskeletal proteins, including actin and beta-catenin.
    • These findings highlight ONOO- as a key mediator in endothelial barrier disruption.