Document Detail


Oxysterol-induced toxicity in R28 and ARPE-19 cells.
MedLine Citation:
PMID:  12718442     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Studies have shown an intimate relationship between cholesterol and retinal diseases; we examined the effects of cholesterol oxides on cultured cells. Using the rat retinal precursor cell line R28 and the human RPE cell line ARPE-19, we investigated the potential cytotoxicity of cholesterol oxides. Cultured R28 and ARPE-19 cells were treated with either 25-hydroxycholesterol and 7-ketocholesterol (0-50 microg/ml). Cell viability was determined by the WST-1 colorimetric assay. Production of reactive oxygen intermediate (ROI) was assessed by a fluorescent probe-based assay (2',7'-dichlorodihydrofluorescein diacetate [H2DCFDA]). To detect the presence of apoptosis, DNA fragmentation gel analysis and Hoescht nuclear staining were performed. Both cholesterol oxides tested were toxic in a time- and dose-dependent fashion to the two cell lines used in this study. Treatment of R28 cells with either 25-hydroxycholesterol or 7-ketocholesterol at a concentration of 25 micro/ml resulted in greater than 50% loss of cell viability after 24 h. ARPE-19 cells were slightly less affected, with a loss of cell viability of approximately 20% and 40% after 24 h-exposure of 25-hydroxycholesterol and 7-ketocholesterol, respectively. DNA fragmentation and chromatin condensation demonstrated apoptotic events occurring in 7-ketocholesterol-treated cells. The fluorescent assay for ROI production showed that after an hour of exposure to 7-ketocholesterol, R28 cells responded with increased levels of ROIs, whereas no immediate production of ROIs were detected with treated ARPE-19 cells. These in vitro findings provide evidence that cholesterol oxides can directly damage cultured retinal and RPE cells. The oxysterol-induced oxidative stress in these cells may be a factor in the pathology of retinal degenerative diseases.
Authors:
John M Ong; Annette M Aoki; Gail M Seigel; Ingrid Sacerio; Raquel Castellon; Anthony B Nesburn; M Cristina Kenney
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Neurochemical research     Volume:  28     ISSN:  0364-3190     ISO Abbreviation:  Neurochem. Res.     Publication Date:  2003 Jun 
Date Detail:
Created Date:  2003-04-29     Completed Date:  2003-07-08     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  7613461     Medline TA:  Neurochem Res     Country:  United States    
Other Details:
Languages:  eng     Pagination:  883-91     Citation Subset:  IM    
Affiliation:
Molecular Eye Research Laboratory, Burns and Allen Research Institute, Cedars-Sinai Medical Center, Los Angeles, California 90048, USA. ongj@cshs.org
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Line
Cell Survival / drug effects*
Dose-Response Relationship, Drug
Humans
Hydroxycholesterols / toxicity*
Hydroxysteroids / toxicity
Ketocholesterols / toxicity*
Kinetics
Rats
Retina
Chemical
Reg. No./Substance:
0/Hydroxycholesterols; 0/Hydroxysteroids; 0/Ketocholesterols; 2140-46-7/25-hydroxycholesterol; 566-28-9/7-ketocholesterol

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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