Oxidative stress causes ERK phosphorylation and cell death in cultured retinal pigment epithelium: prevention of cell death by AG126 and 15-deoxy-delta 12, 14-PGJ2

BMC Ophthalmol. 2003 Mar 21:3:5. doi: 10.1186/1471-2415-3-5.

Abstract

Background: The retina, which is exposed to both sunlight and very high levels of oxygen, is exceptionally rich in polyunsaturated fatty acids, which makes it a favorable environment for the generation of reactive oxygen species. The cytotoxic effects of hydrogen peroxide (H2O2) induced oxidative stress on retinal pigment epithelium were characterized in this study.

Methods: The MTT cell viability assay, Texas-Red phalloidin staining, immunohistochemistry and Western blot analysis were used to assess the effects of oxidative stress on primary human retinal pigment epithelial cell cultures and the ARPE-19 cell line.

Results: The treatment of retinal pigment epithelial cells with H2O2 caused a dose-dependent decrease of cellular viability, which was preceded by a significant cytoskeletal rearrangement, activation of the Extracellular signal-Regulated Kinase, lipid peroxidation and nuclear condensation. This cell death was prevented partially by the prostaglandin derivative, 15d-PGJ2 and by the protein kinase inhibitor, AG126.

Conclusion: 15d-PGJ2 and AG126 may be useful pharmacological tools in the future capable of preventing oxidative stress induced RPE cell death in human ocular diseases.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / metabolism
  • Aldehydes / metabolism
  • Blotting, Western
  • Cell Death
  • Cell Survival / drug effects
  • Cells, Cultured
  • Dose-Response Relationship, Drug
  • Enzyme Inhibitors / pharmacology*
  • Fluorescent Antibody Technique, Indirect
  • Humans
  • Hydrogen Peroxide / toxicity
  • Immunoenzyme Techniques
  • Immunologic Factors / pharmacology*
  • Lipid Peroxidation / drug effects
  • Mitogen-Activated Protein Kinases / antagonists & inhibitors
  • Mitogen-Activated Protein Kinases / metabolism*
  • Oxidative Stress*
  • Phosphorylation
  • Pigment Epithelium of Eye / drug effects*
  • Pigment Epithelium of Eye / metabolism
  • Pigment Epithelium of Eye / pathology
  • Prostaglandin D2 / analogs & derivatives
  • Prostaglandin D2 / pharmacology*
  • Protein-Tyrosine Kinases / antagonists & inhibitors
  • Receptors, Cytoplasmic and Nuclear / agonists
  • Receptors, Cytoplasmic and Nuclear / metabolism
  • Transcription Factors / agonists
  • Transcription Factors / metabolism
  • Tyrphostins / pharmacology*

Substances

  • 15-deoxy-delta(12,14)-prostaglandin J2
  • Actins
  • Aldehydes
  • Enzyme Inhibitors
  • Immunologic Factors
  • Receptors, Cytoplasmic and Nuclear
  • Transcription Factors
  • Tyrphostins
  • AG 127
  • Hydrogen Peroxide
  • Protein-Tyrosine Kinases
  • Mitogen-Activated Protein Kinases
  • 4-hydroxy-2-nonenal
  • Prostaglandin D2