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J. Biol. Chem., Vol. 277, Issue 35, 31929-31937, August 30, 2002
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From the Children's Foundation Research Center at Le Bonheur
Children's Medical Center, University of Tennessee Health Science
Center, Memphis, Tennessee 38103
Apolipoprotein A-IV (apoA-IV) has myriad
functions, including roles as a post-prandial satiety factor and lipid
antioxidant. ApoA-IV is expressed in mammalian small intestine and is
up-regulated in response to lipid absorption. In newborn swine jejunum,
a high fat diet acutely induces a 7-fold increase in apoA-IV
expression. To determine whether apoA-IV plays a role in the transport
of absorbed lipid, swine apoA-IV was overexpressed in a newborn swine enterocyte cell line, IPEC-1, followed by analysis of the expression of
genes related to lipoprotein assembly and lipid transport, as well as
quantitation of lipid synthesis and secretion. A full-length swine apoA-IV cDNA was cloned, sequenced, and inserted into a Vp and Rep gene-deficient adeno-associated
viral vector, containing the cytomegalovirus immediate early
promoter/enhancer and neomycin resistance gene, and was used to
transfect IPEC-1 cells. Control cells were transfected with the same
vector minus the apoA-IV insert. Using neomycin selection,
apoA-IV-overexpressing (+AIV) and control (
Overexpression of Apolipoprotein A-IV Enhances Lipid
Transport in Newborn Swine Intestinal Epithelial Cells*
AIV) clones were isolated
for further study. Both undifferentiated (D) and differentiated (+D)
+AIV cells expressed 40- to 50-fold higher levels of apoA-IV mRNA
and both intracellular and secreted apoA-IV protein compared with AIV
cells. Expression of other genes was not affected by apoA-IV
overexpression in a manner that would contribute to enhanced lipid
secretion. +D +AIV cells secreted 4.9-fold more labeled triacylglycerol
(TG), 4.6-fold more labeled cholesteryl ester (CE), and 2-fold more
labeled phospholipid (PL) as lipoproteins, mostly in the
chylomicron/very low density lipoprotein (VLDL) density range. ApoA-IV
overexpression in IPEC-1 cells enhances basolateral TG, CE, and PL
secretion in chylomicron/VLDL particles. This enhancement is not
associated with up-regulation of other genes involved in lipid
transport. ApoA-IV may play a role in facilitating enterocyte lipid
transport, particularly in the neonate receiving a diet of high fat
breast milk.
*
This work was supported by National Institutes of Health
Grant HD2255 (to D. D. B.) and the Children's Foundation Research Center of Memphis (to D. D. B.).The costs of publication of this article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Children's Foundation
Research Center of Memphis, Le Bonheur Children's Medical Center, Rm.
401, W. Patient Tower, 50 N. Dunlap, Memphis, TN 38103. Tel.:
901-572-5355; Fax: 901-572-4478; E-mail: dblack@utmem.edu.
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