Oxidative stress affects cytoskeletal structure and cell-matrix interactions in cells from an ocular tissue: the trabecular meshwork

J Cell Physiol. 1999 Aug;180(2):182-9. doi: 10.1002/(SICI)1097-4652(199908)180:2<182::AID-JCP6>3.0.CO;2-X.

Abstract

The trabecular meshwork (TM) is a specialized eye tissue that regulates the aqueous humor outflow and controls intraocular pressure. Cells in this tissue are essential for maintenance of the outflow system. Disturbance of the TM cell status by insults such as oxidative stress may lead to elevation of the intraocular pressure and development of glaucoma. In the present study, we investigated the effect of oxidative stress on the adhesion of human TM cells to extracellular matrix (ECM) proteins. Treatment with 1 mM of H2O2 for 10 or 30 min did not affect cell viability, whereas the adhesion of TM cells to fibronectin, laminin, and collagen types I and IV was significantly reduced. Phalloidin and immunostaining also revealed reorganization of actin and vimentin structures. The level of integrins alpha5beta1, alphavbeta3, and beta1 was not altered, although the distribution of paxillin and focal adhesion kinase in focal contacts was reduced. Concomitantly, the level of transcription factor NF-kappaB was enhanced by the H2O2 treatment. Nuclear extracts of the treated cells also contained a heightened NF-kappaB binding activity. These changes persisted for up to 6 h after the H2O2 treatment but were partially recovered by 24 h. We concluded that under sublethal oxidative stress conditions, the TM cell adhesion to the ECM was impaired. The short-term loss of cell-matrix adhesiveness may be related to the rearrangement of cytoskeletal structures. Extensive and repeated oxidative stress in vivo may result in reduced TM cell adhesion, leading to cell loss, compromised TM integrity, and pathologic consequences.

Publication types

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

MeSH terms

  • Actins / analysis
  • Cell Adhesion / physiology
  • Cell Adhesion Molecules / analysis
  • Cells, Cultured
  • Cytoskeletal Proteins / analysis
  • Cytoskeleton / chemistry
  • Cytoskeleton / metabolism*
  • Extracellular Matrix / chemistry
  • Extracellular Matrix / metabolism*
  • Fluorescent Antibody Technique
  • Humans
  • NF-kappa B / analysis
  • NF-kappa B / metabolism
  • Nuclear Proteins / analysis
  • Oxidative Stress / physiology*
  • Paxillin
  • Phosphoproteins / analysis
  • Receptors, Fibronectin / analysis
  • Trabecular Meshwork / chemistry
  • Trabecular Meshwork / cytology*
  • Trabecular Meshwork / metabolism*
  • Vimentin / analysis

Substances

  • Actins
  • Cell Adhesion Molecules
  • Cytoskeletal Proteins
  • NF-kappa B
  • Nuclear Proteins
  • PXN protein, human
  • Paxillin
  • Phosphoproteins
  • Receptors, Fibronectin
  • Vimentin