Crocetin prevents retinal degeneration induced by oxidative and endoplasmic reticulum stresses via inhibition of caspase activity.
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.
Department of Biofunctional Evaluation, Molecular Pharmacology, Gifu Pharmaceutical University, 1-25-4 Daigaku-nishi, Gifu 501-1196, Japan.
Mika
M
Yamauchi
Kazuhiro
K
Tsuruma
Shunsuke
S
Imai
Tomohiro
T
Nakanishi
Naofumi
N
Umigai
Masamitsu
M
Shimazawa
Hideaki
H
Hara
D000284:Administration, Oral; D000818:Animals; D002338:Carotenoids / Q000008:administration & dosage / Q000097:blood / Q000494:pharmacology*; D061945:Caspase Inhibitors; D020169:Caspases / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects / Q000528:radiation effects; D002460:Cell Line; D002839:Chromans / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000187:drug effects* / Q000378:metabolism* / Q000528:radiation effects; D004789:Enzyme Activation / Q000187:drug effects / Q000528:radiation effects; D004791:Enzyme Inhibitors / Q000008:administration & dosage / Q000097:blood / Q000494:pharmacology; D005452:Fluoresceins / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects / Q000528:radiation effects; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects; D008027:Light; D008297:Male; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects / Q000528:radiation effects; D051379:Mice; D010084:Oxidation-Reduction / Q000187:drug effects / Q000528:radiation effects; D018384:Oxidative Stress / Q000187:drug effects* / Q000528:radiation effects; D020419:Photoreceptor Cells, Vertebrate / Q000378:metabolism; D051381:Rats; D012162:Retinal Degeneration / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology / Q000517:prevention & control*; D012165:Retinal Ganglion Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology / Q000528:radiation effects; D014415:Tunicamycin / Q000633:toxicity; D014801:Vitamin A / Q000031:analogs & derivatives; D014966:Xanthenes / Q000378:metabolism
D061945:Caspase Inhibitors; D002839:Chromans; D004791:Enzyme Inhibitors; D005452:Fluoresceins; D014966:Xanthenes; C487773:trans-sodium crocetinate; C065013:2',7'-dichlorodihydrofluorescein; D014415:Tunicamycin; D014801:Vitamin A; D002338:Carotenoids; D006861:Hydrogen Peroxide; D020169:Caspases; C010643:6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid