Crocetin prevents retinal degeneration induced by oxidative and endoplasmic reticulum stresses via inhibition of caspase activity

Eur J Pharmacol. 2011 Jan 10;650(1):110-9. doi: 10.1016/j.ejphar.2010.09.081. Epub 2010 Oct 14.

Abstract

Crocetin is a carotenoid that is the aglicone of crocin, which are found in saffron stigmas (Crocus sativus L.) and gardenia fruit (Gardenia jasminoides Ellis). In this study, we investigated the effects of crocetin on retinal damage. To examine whether crocetin affects stress pathways, we investigated intracellular oxidation induced by reactive oxygen species, expression of endoplasmic reticulum (ER) stress-related proteins, disruption of the mitochondrial membrane potential (ΔΨ(m)), and caspases activation. In vitro, we employed cultured retinal ganglion cells (RGC-5, a mouse ganglion cell-line transformed using E1A virus). Cell damage was induced by tunicamycin or hydrogen peroxide (H(2)O(2)) exposure. Crocetin at a concentration of 3μM showed the inhibitory effect of 50-60% against tunicamycin- and H(2)O(2)-induced cell death and inhibited increase in caspase-3 and -9 activity. Moreover, crocetin inhibited the enzymatic activity of caspase-9 in a cell-free system. In vivo, retinal damage in mice was induced by exposure to white light at 8000lx for 3h after dark adaptation. Photoreceptor damage was evaluated by measuring the outer nuclear layer thickness at 5days after light exposure and recording the electroretinogram (ERG). Retinal cell damage was also detected with Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining at 48h after light exposure. Crocetin at 100mg/kg, p.o. significantly inhibited photoreceptor degeneration and retinal dysfunction and halved the expression of TUNEL-positive cells. These results indicate that crocetin has protective effects against retinal damage in vitro and in vivo, suggesting that the mechanism may inhibit increase in caspase-3 and -9 activities after retinal damage.

MeSH terms

  • Administration, Oral
  • Animals
  • Carotenoids / administration & dosage
  • Carotenoids / blood
  • Carotenoids / pharmacology*
  • Caspase Inhibitors*
  • Caspases / metabolism
  • Cell Death / drug effects
  • Cell Death / radiation effects
  • Cell Line
  • Chromans / pharmacology
  • Endoplasmic Reticulum / drug effects*
  • Endoplasmic Reticulum / metabolism*
  • Endoplasmic Reticulum / radiation effects
  • Enzyme Activation / drug effects
  • Enzyme Activation / radiation effects
  • Enzyme Inhibitors / administration & dosage
  • Enzyme Inhibitors / blood
  • Enzyme Inhibitors / pharmacology
  • Fluoresceins / metabolism
  • Gene Expression Regulation / drug effects
  • Gene Expression Regulation / radiation effects
  • Humans
  • Hydrogen Peroxide / toxicity
  • Intracellular Space / drug effects
  • Intracellular Space / metabolism
  • Intracellular Space / radiation effects
  • Light
  • Male
  • Membrane Potential, Mitochondrial / drug effects
  • Membrane Potential, Mitochondrial / radiation effects
  • Mice
  • Oxidation-Reduction / drug effects
  • Oxidation-Reduction / radiation effects
  • Oxidative Stress / drug effects*
  • Oxidative Stress / radiation effects
  • Photoreceptor Cells, Vertebrate / metabolism
  • Rats
  • Retinal Degeneration / enzymology
  • Retinal Degeneration / metabolism*
  • Retinal Degeneration / pathology
  • Retinal Degeneration / prevention & control*
  • Retinal Ganglion Cells / drug effects
  • Retinal Ganglion Cells / metabolism
  • Retinal Ganglion Cells / pathology
  • Retinal Ganglion Cells / radiation effects
  • Tunicamycin / toxicity
  • Xanthenes / metabolism

Substances

  • Caspase Inhibitors
  • Chromans
  • Enzyme Inhibitors
  • Fluoresceins
  • Xanthenes
  • 2',7'-dichlorodihydrofluorescein
  • Tunicamycin
  • crocetin
  • Carotenoids
  • Hydrogen Peroxide
  • Caspases
  • 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid