H(2)O(2) inhibits alveolar epithelial wound repair in vitro by induction of apoptosis

Am J Physiol Lung Cell Mol Physiol. 2004 Aug;287(2):L448-53. doi: 10.1152/ajplung.00177.2003. Epub 2004 Apr 30.

Abstract

Reactive oxygen species (ROS) are released into the alveolar space and contribute to alveolar epithelial damage in patients with acute lung injury. However, the role of ROS in alveolar repair is not known. We studied the effect of ROS in our in vitro wound healing model using either human A549 alveolar epithelial cells or primary distal lung epithelial cells. We found that H(2)O(2) inhibited alveolar epithelial repair in a concentration-dependent manner. At similar concentrations, H(2)O(2) also induced apoptosis, an effect seen particularly at the edge of the wound, leading us to hypothesize that apoptosis contributes to H(2)O(2)-induced inhibition of wound repair. To learn the role of apoptosis, we blocked caspases with the pan-caspase inhibitor N-benzyloxycarbonyl-Val-Ala-Asp (zVAD). In the presence of H(2)O(2), zVAD inhibited apoptosis, particularly at the wound edge and, most importantly, maintained alveolar epithelial wound repair. In H(2)O(2)-exposed cells, zVAD also maintained cell viability as judged by improved cell spreading and/or migration at the wound edge and by a more normal mitochondrial potential difference compared with cells not treated with zVAD. In conclusion, H(2)O(2) inhibits alveolar epithelial wound repair in large part by induction of apoptosis. Inhibition of apoptosis can maintain wound repair and cell viability in the face of ROS. Inhibiting apoptosis may be a promising new approach to improve repair of the alveolar epithelium in patients with acute lung injury.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Chloromethyl Ketones / pharmacology
  • Apoptosis / drug effects*
  • Caspase Inhibitors
  • Cell Line
  • Cell Movement / drug effects
  • Cysteine Proteinase Inhibitors / pharmacology
  • Humans
  • Hydrogen Peroxide / pharmacology*
  • In Vitro Techniques
  • Oxidants / pharmacology*
  • Pulmonary Alveoli / cytology*
  • Pulmonary Alveoli / drug effects
  • Respiratory Mucosa / cytology*
  • Respiratory Mucosa / drug effects
  • Wound Healing / drug effects*

Substances

  • Amino Acid Chloromethyl Ketones
  • Caspase Inhibitors
  • Cysteine Proteinase Inhibitors
  • Oxidants
  • benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone
  • Hydrogen Peroxide