Effect of chronic incubation of CaCo-2 cells with eicosapentaenoic acid (20:5, <i>n</i>-3) and oleic acid (18:1, <i>n</i>-9) on triacylglycerol production

Ranheim, Trine; Gedde-Dahl, Ane; Rustan, Arild C.; Drevon, Christian A.

doi:10.1042/bj3030155

Cited by 25 publications

(18 citation statements)

References 32 publications

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“…We have shown that secretion of lipoproteins can be further modulated by OA supplementation to obtain secretion of small and large CM. In some studies, OA was delivered to Caco-2 cells as part of TC micelles (21,32,50,51). In these studies the concentrations of OA used were below Ϸ0.8 mM; a concentration that we found to be less than the optimum amount required for CM secretion (Fig.…”

Section: Discussionmentioning

confidence: 99%

Assembly and Secretion of Chylomicrons by Differentiated Caco-2 Cells

Luchoomun¹,

Hussain

1999

Journal of Biological Chemistry

158

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To develop a cell culture model for chyclomicron (CM) assembly, the apical media of differentiated Caco-2 cells were supplemented with oleic acid (OA) together with either albumin or taurocholate (TC). The basolateral media were subjected to sequential density gradient ultracentrifugations to obtain large CM, small CM, and very low density lipoproteins (VLDL), and the distribution of apoB in these fractions was quantified. In the absence of OA, apoB was secreted as VLDL/LDL size particles. Addition of OA (>0.8 mM) with TC, but not with albumin, resulted in the secretion of one-third of apoB as CM. Lipid analysis revealed that half of the secreted phospholipids (PL) and triglycerides (TG) were associated with CM. In CM, TG were 7-11-fold higher than PL indicating that CM were TG-rich particles. Secreted CM contained apoB100, apoB48, and other apolipoproteins. Secretion of large CM was specifically inhibited by Pluronic L81, a detergent known to inhibit CM secretion in animals. These studies demonstrate that differentiated Caco-2 cells assemble and secrete CM in a manner similar to enterocytes in vivo. Next, experiments were performed to identify the sources of lipids used for lipoprotein assembly. Cells were labeled with [ 3 H]glycerol for 12 h, washed, and supplemented with OA, TC, and [ 14 C] glycerol for various times to induce CM assembly and to radiolabel nascent lipids. TG and PL were extracted from cells and media and the association of preformed and nascent lipids with lipoproteins was determined. All the lipoproteins contained higher amounts of preformed PL compared with nascent PL. VLDL contained equal amounts of nascent and preformed TG, whereas CM contained higher amounts of nascent TG even when nascent TG constituted a small fraction of the total cellular pool. These studies indicate that nascent TG and preformed PL are preferentially used for CM assembly and provide a molecular explanation for the in vivo observations that the fatty acid composition of TG, but not PL, of secreted CM reflects the composition of dietary fat. It is proposed that in the intestinal cells the preformed PL from the endoplasmic reticulum bud off with apoB as primordial particles and the assembly of larger lipoproteins is dependent on the synthesis and delivery of nascent TG to these particles.

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Section: Discussionmentioning

confidence: 99%

Assembly and Secretion of Chylomicrons by Differentiated Caco-2 Cells

Luchoomun¹,

Hussain

1999

Journal of Biological Chemistry

158

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“…Also, studies with Caco-2 cells have demonstrated that incubation with (n-3) VLCPUFAs lowered triglyceride synthesis and secretion (11)(12)(13)(14)(15). Finally, a single study with cultured intestinal biopsies obtained from malnourished patients demonstrated a decreased triglyceride resynthesis in cells from subjects that had consumed an (n-3) VLCPUFA supplemented diet for 2 weeks (30).…”

Section: Discussionmentioning

confidence: 95%

“…Altered enterocyte function and increased chylomicron clearance from plasma could contribute to the observed effects of fish oil (6,9,10). Some in vitro studies have suggested that (n-3) VLCPUFAs lower triglyceride resynthesis and chylomicron/ triglyceride secretion from enterocytes (11)(12)(13)(14)(15). However, chylomicron secretion appeared not to be decreased in rats fed (n-3) VLCPUFAs (8,16).…”

mentioning

confidence: 91%

Fish Oil Feeding is Associated with an Increased Accumulation of Dietary Lipids in Enterocytes: Results from an in vivo Study in Rats

Larsen

Marckmann

Hansen³

et al. 2003

Scandinavian Journal of Gastroenterology

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“…However, the overall increased secretion could also result from either increased synthesis due to enhanced transcription or due to decreased posttranslational degradation. For example, the increase in apoB secretion following addition of oleic acid plus taurocholate [34] to Caco-2 cultures is not due to increased synthesis, but due to reduced degradation [11,13,46], while the increased apoB secretion following stimulation with transforming growth factor β has been shown to result from increased transcription of the apoB gene [41,50]. Further experiments are required to determine if reduced degradation and/or increased transcription is responsible for the increased apoB secretion due to growth of Caco-2 cells on HACT.…”

Section: Discussionmentioning

confidence: 99%

Fibrillar collagen type I stimulation of apolipoprotein B secretion in Caco-2 cells is mediated by β1 integrin

Ratcliffe

Iqbal

Hussain

et al. 2009

Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biolog

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Caco-2 cells spontaneously differentiate into enterocyte-like cells and secrete apolipoprotein B (apoB) lipoproteins. We evaluated the effect of different extracellular matrix proteins on lipoprotein secretion by these cells. Caco-2 cells grown on human amnion connective tissue (HACT) secreted twice as much apoB as control cells on Transwells, but secreted similar amounts of apoA1. Cells cultured on fibrillar collagen type I secreted increased amounts of apoB similar to the cells cultured on HACT, but cells cultured on non-fibrillar collagen type I, type IV collagen or laminin-1 did not. The increased secretion was nullified by a function inhibiting anti-integrin β1 monoclonal antibody. Therefore, interactions between type I collagen and β1 integrins augment apoB secretion by Caco-2 cells. Cells on HACT formed a more uniform columnar epithelium with lipid droplets polarized to the basolateral membrane. We also studied the effect of extracellular matrix proteins on transepithelial resistance (TER) of differentiated Caco-2 cells. TER in cells cultured on HACT was similar to that on Transwells, but cells on laminin-1 and collagen IV exhibited higher TER. Thus, various extracellular matrix proteins regulate apoB secretion and TER differently. This new observation that extracellular matrix proteins can enhance apoB secretion in Caco-2 cells could be useful to explore the modulation of lipid transport by these proteins.

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Effect of chronic incubation of CaCo-2 cells with eicosapentaenoic acid (20:5, n-3) and oleic acid (18:1, n-9) on triacylglycerol production

Cited by 25 publications

References 32 publications

Assembly and Secretion of Chylomicrons by Differentiated Caco-2 Cells

Assembly and Secretion of Chylomicrons by Differentiated Caco-2 Cells

Fish Oil Feeding is Associated with an Increased Accumulation of Dietary Lipids in Enterocytes: Results from an in vivo Study in Rats

Fibrillar collagen type I stimulation of apolipoprotein B secretion in Caco-2 cells is mediated by β1 integrin

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