Epigallocatechin gallate (EGCG) prevents H2O2-induced oxidative stress in primary rat retinal pigment epithelial cells

Curr Eye Res. 2014 Sep;39(9):944-52. doi: 10.3109/02713683.2014.885532. Epub 2014 Feb 21.

Abstract

Purpose: To determine whether the green tea polyphenol epigallocatechin gallate (EGCG) could prevent H(2)O(2)-induced oxidative stress in primary rat retinal pigment epithelial cells.

Methods: Primary cultures of retinal pigment epithelium (RPE) cells were established from Long-Evans newborn rats. RPE cells were pretreated with various concentrations of EGCG for 24 h before being exposed to hydrogen peroxide (H(2)O(2)) for 2 h to induce oxidative stress. Cell metabolic activity was measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cell death was quantified by flow cytometry using propidium iodide (PI).

Results: Treatment of RPE cells with EGCG alone does not affect the cell viability up to 50 µM. Exposure of RPE cells to 600 µM H(2)O(2) caused a significant decrease in cell viability; whereas pretreatment with 10, 25, and 50 µM EGCG significantly reduced this decrease in a dose-dependent manner. The proportion of PI-positive cells increased significantly in cultures treated with H(2)O(2) alone; whereas pretreatment of RPE cells with 50 µM EGCG significantly reduced H(2)O(2)-induced RPE cell death.

Conclusions: Our study shows that EGCG pretreatment can protect primary rat RPE cells from H(2)O(2)-induced death. This suggests potential effect of EGCG in the prevention of retinal diseases associated with H(2)O(2)-induced oxidative stress.

Keywords: Age-related macular degeneration; epigallocatechin gallate; hydrogen peroxide; oxidative stress; primary retinal pigment epithelial cell.

MeSH terms

  • Animals
  • Animals, Newborn
  • Antioxidants / pharmacology*
  • Apoptosis
  • Catechin / analogs & derivatives*
  • Catechin / pharmacology
  • Cell Survival / drug effects
  • Cells, Cultured
  • Dose-Response Relationship, Drug
  • Flow Cytometry
  • Hydrogen Peroxide / toxicity*
  • Oxidants / toxicity*
  • Oxidative Stress / drug effects*
  • Rats
  • Rats, Long-Evans
  • Retinal Pigment Epithelium / drug effects*
  • Retinal Pigment Epithelium / metabolism
  • Retinal Pigment Epithelium / pathology

Substances

  • Antioxidants
  • Oxidants
  • Catechin
  • Hydrogen Peroxide
  • epigallocatechin gallate