Caspase-3 and its inhibitor Ac-DEVD-CHO in rat lens epithelial cell apoptosis induced by hydrogen in vitro

Chin Med J (Engl). 2003 Jul;116(7):1034-8.

Abstract

Objective: To investigate the role of caspase-3 and its inhibitor Ac-DEVD-CHO in rat lens epithelial cell apoptosis induced by hydrogen peroxide (H(2)O(2)) in vitro.

Methods: Rat lenses were incubated in modified Eagle's medium containing 2 mmol/L H(2)O(2) to induce apoptosis in vitro. Apoptosis in lens epithelial cells was assessed by transmission electron microscopy and annexin V-propidium iodide (PI) double staining flow cytometry after 12, 24 and 48 h of incubation. The activity of caspase-3 was analyzed by western blotting.

Results: Observations under transmission electron microscopy revealed that 2 mmol/L H(2)O(2) could effectively induce lens epithelial cell apoptosis in vitro. Caspase-3 activity increased during cell apoptosis and the peak measurement occurred at 24 h after treatment with H(2)O(2). Cell apoptosis was blocked by caspase-3 inhibitor Ac-DEVD-CHO.

Conclusions: The activation of caspase-3 plays an important role in executing apoptosis in H(2)O(2)-treated lens epithelial cells and in the formation of cataract. The caspase-3 inhibitor Ac-DEVD-CHO may effectively prevent lens epithelial cell apoptosis caused by oxidative injury.

MeSH terms

  • Animals
  • Apoptosis / drug effects*
  • Caspase 3
  • Caspase Inhibitors*
  • Caspases / metabolism
  • Caspases / physiology*
  • Cysteine Proteinase Inhibitors / pharmacology*
  • Enzyme Activation
  • Epithelial Cells / cytology
  • Female
  • Hydrogen Peroxide / pharmacology*
  • Lens, Crystalline / cytology*
  • Oligopeptides / pharmacology*
  • Organ Culture Techniques
  • Rats
  • Rats, Sprague-Dawley

Substances

  • Caspase Inhibitors
  • Cysteine Proteinase Inhibitors
  • Oligopeptides
  • acetyl-aspartyl-glutamyl-valyl-aspartal
  • Hydrogen Peroxide
  • Casp3 protein, rat
  • Caspase 3
  • Caspases