Apolipoprotein B100 secretion by cultured ARPE-19 cells is modulated by alteration of cholesterol levels

Tinghuai Wu, Masashi Fujihara, Jane Tian, Miroslava Jovanovic, Celene Grayson, Marisol Cano, Peter Gehlbach, Philippe Margaron, James Handa

Research output: Contribution to journalArticle

Abstract

Cholesteryl ester rich apolipoprotein B100 (apoB100) lipoproteins accumulate in Bruch's membrane before the development of age-related macular degeneration. It is not known if these lipoproteins come from the circulation or local ocular tissue. Emerging, but incomplete evidence suggests that the retinal pigmented epithelium (RPE) can secrete lipoproteins. The purpose of this investigation was to determine (i) whether human RPE cells synthesize and secrete apoB100, and (ii) whether this secretion is driven by cellular cholesterol, and if so, (iii) whether statins inhibit this response. The established, human derived ARPE-19 cells challenged with 0-0.8 mM oleic acid accumulated cellular cholesterol, but not triglycerides. Oleic acid increased the amount of apoB100 protein recovered from the medium by both western blot analysis and 35S-radiolabeled immunoprecipitation while negative stain electron microscopy showed lipoprotein-like particles. Of nine statins evaluated, lipophilic statins induced HMG-CoA reductase mRNA expression the most. The lipophilic Cerivastatin (5 μM) reduced cellular cholesterol by 39% and abrogated apoB100 secretion by 3-fold. In contrast, the hydrophilic statin Pravastatin had minimal effect on apoB100 secretion. These data suggest that ARPE-19 cells synthesize and secrete apoB100 lipoproteins, that this secretion is driven by cellular cholesterol, and that statins can inhibit apoB100 secretion by reducing cellular cholesterol.

Original languageEnglish (US)
Pages (from-to)1734-1744
Number of pages11
JournalJournal of Neurochemistry
Volume114
Issue number6
DOIs
StatePublished - 2010

Fingerprint

Apolipoproteins
Hydroxymethylglutaryl-CoA Reductase Inhibitors
Cholesterol
Lipoproteins
Oleic Acid
Epithelium
Bruch Membrane
Hydroxymethylglutaryl CoA Reductases
Pravastatin
Cholesterol Esters
Macular Degeneration
Immunoprecipitation
Electron microscopy
Electron Microscopy
Triglycerides
Coloring Agents
Western Blotting
Tissue
Membranes
Messenger RNA

Keywords

  • age-related macular degeneration
  • apolipoprotein B100
  • basal deposits
  • Bruch's membrane
  • drusen
  • retinal pigmented epithelium

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience
  • Medicine(all)

Cite this

Apolipoprotein B100 secretion by cultured ARPE-19 cells is modulated by alteration of cholesterol levels. / Wu, Tinghuai; Fujihara, Masashi; Tian, Jane; Jovanovic, Miroslava; Grayson, Celene; Cano, Marisol; Gehlbach, Peter; Margaron, Philippe; Handa, James.

In: Journal of Neurochemistry, Vol. 114, No. 6, 2010, p. 1734-1744.

Research output: Contribution to journalArticle

Wu, Tinghuai ; Fujihara, Masashi ; Tian, Jane ; Jovanovic, Miroslava ; Grayson, Celene ; Cano, Marisol ; Gehlbach, Peter ; Margaron, Philippe ; Handa, James. / Apolipoprotein B100 secretion by cultured ARPE-19 cells is modulated by alteration of cholesterol levels. In: Journal of Neurochemistry. 2010 ; Vol. 114, No. 6. pp. 1734-1744.
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