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Originally published In Press as doi:10.1194/jlr.M700263-JLR200 on August 2, 2007 Originally published In Press as doi:10.1194/jlr.M700263-JLR200 on July 20, 2007

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Journal of Lipid Research, Vol. 48, 2283-2294, October 2007
Copyright © 2007 by American Society for Biochemistry and Molecular Biology

Mechanisms of provitamin A (carotenoid) and vitamin A (retinol) transport into and out of intestinal Caco-2 cells

Alexandrine During1,* and Earl H. Harrison{dagger}

* Department of International Health, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD 21205
{dagger} Department of Human Nutrition, Ohio State University, Columbus, OH 43210

Published, JLR Papers in Press, August 2, 2007.

1 To whom correspondence should be addressed. e-mail: during{at}bioc.ucl.ac.be

The purpose of this study was to compare the mechanisms of intestinal retinol (ROL) and carotenoid transport. When differentiated Caco-2 cells were incubated with ROL for varying times, cellular ROL plateaued within 2 h, whereas retinyl ester (RE) formation increased continuously. ROL and RE efflux into basolateral medium (BM) increased linearly with time, ROL in the nonlipoprotein fraction and REs in chylomicrons (CMs). In contrast to carotenoids, ROL uptake was proportional to ROL concentration (0.5–110 µM). ROL efflux into BM occurred via two processes: a) a saturable process at low concentrations (<10 µM) and b) a nonsaturable process at higher concentrations. When ROL-loaded cells were maintained on retinoid-free medium, free ROL, but not REs, was secreted into BM. Glyburide significantly reduced ROL efflux but not ROL uptake. Inhibition of ABCA1 protein expression by small interfering RNAs decreased ROL efflux but not carotenoid efflux. Scavenger receptor class B type I (SR-BI) inhibition did not affect ROL transport but decreased carotenoid uptake. The present data suggest that a) ROL enters intestinal cells by diffusion, b) ROL efflux is partly facilitated, probably by the basolateral transporter ABCA1, and c) newly synthesized REs, but not preformed esters, are incorporated into CM and secreted. In contrast to ROL transport, carotenoid uptake is mediated by the apical transporter SR-BI, and carotenoid efflux occurs exclusively via their secretion in CM.

Supplementary key words retinoids • chylomicrons • glyburide • ATP binding cassette, subfamily A, member 1 • scavenger receptor class B type I • Niemann-Pick disease type C1 gene-like 1

Abbreviations: ABCA1, ATP-binding cassette, sub-family A, member 1; AM, apical medium; apoB, apolipoprotein B; BM, basolateral medium; CM, chylomicron; LP2000, LipofectamineTM 2000; NPC1L1, Niemann-Pick disease type C1 gene-like 1; OA, oleic acid; RE, retinyl ester; ROL, retinol; siRNA or RNAi, small interfering RNA; SR-BI, scavenger receptor class B type I; TC, taurocholate; TG, triglyceride


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