J. Lipid Res. Acyl Labeled PIP's available August 1, 2008
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Journal of Lipid Research, Vol 38, 2273-2280, Copyright © 1997 by Lipid Research, Inc.

ARTICLES

Mechanism of the slow induction of apolipoprotein A-I synthesis by retinoids in cynomolgus hepatocytes: involvement of retinoic acid and retinoid X receptors

A Kaptein, DM Neele, J Twisk, HF Hendriks, T Kooistra and HM Princen
Gaubius Laboratory, TNO-PG, Leiden, The Netherlands.

We showed previously that retinoids stimulate apolipoprotein A-I (apoA- I) synthesis in cultured cynomolgus hepatocytes only after a 24-h lag phase. Here we report on the biochemical background of the slow response, the requirement for high retinoic acid concentrations, and the involvement of different retinoid receptors. The time course of the effect of 10 microM all-trans retinoic acid (at-RA) on apoA-I mRNA levels and protein secretion were comparable, i.e., minor increases were observed after a 24-h incubation and mRNA levels were increased 2.2- and 3.5-fold after 48 h and 72 h, respectively. In contrast, apoA- I gene transcription was already increased (2.6-fold) after a 4-h incubation with 10 microM at-RA. At-RA disappeared rapidly from the cultures: after 2 h of incubation 40% of the added amount was left and after 24 h only 2%. RAR beta mRNA and gene expression were increased after incubation with 10 microM at-RA, whereas RAR alpha and RXR alpha mRNA levels and expression remained unchanged. No transcriptional activity and mRNA for other retinoid receptors were detectable. Both RAR-selective (TTNPB) and RXR-selective (3-methyl-TTNEB) agonists induced apoA-I synthesis at 1 and 10 microM. These results show that i) the slow increase in apoA-I secretion is caused by a slow increase of its mRNA level; ii) the apoA-I gene transcription in cynomolgus hepatocytes is induced rapidly by retinoids; iii) the added at-RA disappeared rapidly from the cultures, explaining the necessity for high initial concentrations; iv) RLR alpha and/or RAR beta and RXR alpha are involved in the activation of apoA-I expression by retinoids.
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