Abstract:
PURPOSE:To characterize the possible cytotoxic effects of oxysterols (7beta-hydroxycholesterol (7beta-OH), 25-hydroxycholesterol (25-OH)) in human retinal pigment epithelial cells (ARPE-19) and to detail the relationships between some of these effects. METHODS:ARPE-19 cells were treated with 7beta-OH and 25-OH. Cell viability was measured with the MTT assay. Membrane permeability, mitochondrial potential, and lysosomal integrity were measured by flow cytometry with propidium iodide, DiOC6(3), and acridine orange, respectively. Cell death was characterized by staining with Hoechst 33342, transmission electron microscopy, and analysis of the DNA fragmentation pattern. Caspase activity was examined with fluorochrome-labeled inhibitors of caspases (FLICA) and Western blotting. Immunofluorescence staining was used to visualize the cellular distribution of cytochrome c (Cyt-c) and apoptosis-inducing factor (AIF). The effect of the cathepsin inhibitor (z-FA-fmk) on oxysterol-induced cell death was evaluated. RESULTS:Cell viability of ARPE-19 cells was decreased with 7beta-OH, whereas 25-OH had no cytotoxic effects. Loss of mitochondrial potential and lysosomal destabilization was associated with 7beta-OH-induced cell death, few morphologically apoptotic cells were identified, and no internucleosomal DNA fragmentation was found. Slight caspase activation was detected with FLICA, and no caspase-3 activation was revealed. A pronounced relocalization of Cyt-c and AIF was observed. Noteworthy, z-FA-fmk was able to prevent cell death. CONCLUSION:7beta-OH induced a caspase-3-independent mode of cell death associated with lysosomal destabilization, which could play a key role in the signaling pathways leading to cell death, as shown by the ability of z-FA-fmk to counteract the cytotoxic effects of 7beta-OH.
journal_name
Curr Eye Resjournal_title
Current eye researchauthors
Malvitte L,Montange T,Vejux A,Joffre C,Bron A,Creuzot-Garcher C,Lizard Gdoi
10.1080/02713680802337397subject
Has Abstractpub_date
2008-09-01 00:00:00pages
769-81issue
9eissn
0271-3683issn
1460-2202pii
902507057journal_volume
33pub_type
杂志文章abstract::Introduction: Metabolic syndrome is a disorder characterized by a constellation of findings including truncal obesity, elevated blood pressure, abnormal cholesterol levels, and high blood glucose. Recent evidence suggests that metabolic syndrome may be associated with increased risk of age-related macular degeneration...
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更新日期:1987-07-01 00:00:00
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更新日期:1987-08-01 00:00:00
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pub_type: 临床试验,杂志文章,多中心研究,随机对照试验
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