Oxidized low density lipoprotein-induced inhibition of processing of photoreceptor outer segments by RPE.
Invest Ophthalmol Vis Sci
; 42(11): 2714-20, 2001 Oct.
Article
em En
| MEDLINE
| ID: mdl-11581220
ABSTRACT
PURPOSE:
To examine the effects of oxidized low-density lipoproteins (oxLDL) on phagocytosis and processing of photoreceptor outer segments (OS) by retinal pigment epithelial (RPE) cells.METHODS:
Confluent cultures of RPE-J cells were pretreated with oxLDL or LDL, and the effects of such treatment on the processing of added OS was determined. Processing was determined either by the degradation of 125I-labeled OS to trichloroacetic acid-soluble label or by the cleavage of rhodopsin observed on Western blot analysis of cell lysates separated by sucrose density gradient fractionation. Binding to and uptake of OS by RPE-J cells was assessed by determining the fluorescence of FITC-labeled OS before and after quenching with trypan blue.RESULTS:
OxLDL induced a significant decrease in the degradation of 125I-OS in RPE-J cells but no reductions in either binding or uptake, when a 24-hour recovery period was inserted between treatment with oxLDL and challenge with OS. Rhodopsin cleavage increased in a time-dependent manner after phagocytosis of OS by RPE-J cells. The small guanosine triphosphatase (GTPase), Rab5, was first found in phagosome fractions containing rhodopsin and its cleavage products, and at later times of challenge, in more dense fractions representing phagolysosomes. These were assessed by the colocalization of rhodopsin cleavage products in density fractions with cathepsin D, a marker of lysosomes. OxLDL induced a reduction in rhodopsin cleavage product formation and in phagosome-lysosome fusion (maturation). It also reduced the time-dependent shift of rhodopsin to higher density fractions containing more cathepsin D without any detectable reduction in the expression of cathepsin D or in acid protease activity.CONCLUSIONS:
OxLDL induces a reduction in the processing of OS by RPE by perturbing the fusion of lysosomes with phagosomes.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Fagocitose
/
Epitélio Pigmentado Ocular
/
Segmento Externo da Célula Bastonete
/
Lipoproteínas LDL
Limite:
Animals
Idioma:
En
Revista:
Invest Ophthalmol Vis Sci
Ano de publicação:
2001
Tipo de documento:
Article
País de afiliação:
Estados Unidos