Lutein transport by Caco-2 TC-7 cells occurs partly by a facilitated process involving the scavenger receptor class B type I (SR-BI)

Author:

REBOUL Emmanuelle1234,ABOU Lydia5,MIKAIL Céline5,GHIRINGHELLI Odette1234,ANDRÉ Marc1234,PORTUGAL Henri5,JOURDHEUIL-RAHMANI Dominique12346,AMIOT Marie-Josèphe1234,LAIRON Denis1234,BOREL Patrick1234

Affiliation:

1. UMR 476 INSERM, Faculté de Médecine, 27 Bd Jean Moulin, 13385 Marseille Cedex 5, France

2. 1260 INRA, Marseille F-13385, France

3. Univ Aix-Marseille, Marseille F-13385, France

4. IPHM, Marseille F-13385, France

5. Service de Chimie Analytique, Faculté de Pharmacie, 27 Bd Jean Moulin, Marseille 13005, France

6. Laboratoire de Biochimie et Sémiologie Cliniques, Faculté de Pharmacie, 27 Bd Jean Moulin, Marseille, 13005, France

Abstract

The carotenoid lutein is thought to play a role in the human eye and to protect against age-related macular degeneration. Lutein transport in the human intestine has not been characterized. We examined lutein transport processes using Caco-2 TC-7 monolayers as a model for human intestinal epithelium. Purified lutein was mixed with phospholipids, lysophospholipids, cholesterol, mono-olein, oleic acid and taurocholate to obtain lutein-rich mixed micelles that mimicked those found under physiological conditions. The micelles were added to the apical side of Caco-2 TC-7 cell monolayers for 30 min or 3 h at 37 °C. Absorbed lutein, i.e. the sum of lutein recovered in the scraped cells and in the basolateral chamber, was quantified by HPLC. Transport rate was measured (i) as a function of time (from 15 to 60 min), (ii) as a function of micellar lutein concentration (from 1.5 to 15 μM), (iii) at 4 °C, (iv) in the basolateral to apical direction, (v) after trypsin pretreatment, (vi) in the presence of β-carotene and/or lycopene, (vii) in the presence of increasing concentrations of antibody against SR-BI (scavenger receptor class B type 1) and (viii) in the presence of increasing concentrations of a chemical inhibitor of the selective transfer of lipids mediated by SR-BI, i.e. BLT1 (blocks lipid transport 1). The rate of transport of lutein as a function of time and as a function of concentration was saturable. It was significantly lower at 4 °C than at 37 °C (approx. 50%), in the basal to apical direction than in the opposite direction (approx. 85%), and after trypsin pretreatment (up to 45%). Co-incubation with β-carotene, but not lycopene, decreased the lutein absorption rate (approx. 20%) significantly. Anti-SR-BI antibody and BLT1 significantly impaired the absorption rate (approx. 30% and 57% respectively). Overall, these results indicate that lutein absorption is, at least partly, protein-mediated and that some lutein is taken up through SR-BI.

Publisher

Portland Press Ltd.

Subject

Cell Biology,Molecular Biology,Biochemistry

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