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Oxysterol-induced toxicity in R28 and ARPE-19 cells
John M Ong1, Annette M Aoki, Gail M Seigel
1Molecular Eye Research Laboratory, Burns and Allen Research Institute, Cedars-Sinai Medical Center, Los Angeles, California 90048, USA. ongj@cshs.org
Neurochemical Research
|April 30, 2003
Summary
Cholesterol oxides are toxic to retinal cells, causing cell death and oxidative stress. These findings suggest a role for oxysterols in the development of retinal degenerative diseases.
Area of Science:
- Ophthalmology
- Cell Biology
- Toxicology
Background:
- Cholesterol's link to retinal diseases is established.
- Cholesterol oxides (oxysterols) are potential contributors to retinal pathology.
Purpose of the Study:
- To investigate the cytotoxic effects of cholesterol oxides on cultured retinal cells.
- To determine if cholesterol oxides induce oxidative stress and apoptosis in retinal cells.
Main Methods:
- Utilized rat retinal precursor (R28) and human RPE (ARPE-19) cell lines.
- Treated cells with 25-hydroxycholesterol and 7-ketocholesterol.
- Assessed cell viability (WST-1), reactive oxygen intermediate (ROI) production (H2DCFDA), and apoptosis (DNA fragmentation, Hoescht staining).
Main Results:
- Both cholesterol oxides exhibited dose- and time-dependent toxicity to R28 and ARPE-19 cells.
- Significant cell viability loss (>50% in R28 cells) observed at 25 microg/ml.
- 7-ketocholesterol induced apoptosis and increased ROI production in R28 cells.
Conclusions:
- Cholesterol oxides directly damage cultured retinal and RPE cells.
- Oxysterol-induced oxidative stress may contribute to retinal degenerative disease pathology.