van Greevenbroek M M, Voorhout W F, Erkelens D W, van Meer G, de Bruin T W
Department of Medicine, Faculty of Medicine, Utrecht University, The Netherlands.
J Lipid Res. 1995 Jan;36(1):13-24.
Polarized monolayers of intestinal Caco-2 cells were used to study the effects of saturated palmitic acid (16:0) and polyunsaturated linoleic acid (18:2) on triglyceride synthesis and lipoprotein secretion. Monolayers were incubated for 24 h, at the apical or lumenal side, with palmitic acid (16:0) or linoleic acid (18:2) in physiological concentrations. Incubation with 1.0 mM 16:0 or 18:2 resulted in differences in the composition and amount of secreted lipoproteins. Radiolabeled lipids in the lipoproteins secreted during incubation with 18:2 were found in the chylomicron/VLDL (very low density lipoprotein) density whereas with 16:0 the secreted lipoproteins were in the intermediate density/low density lipoprotein (IDL/LDL) density range. More triglyceride was secreted into the (basolateral) medium during incubation with 1.0 mM 18:2 (41 +/- 12% of total triglyceride synthesized) than with 1.0 mM 16:0 (18 +/- 3% of total). The biochemical findings correlate with conspicuous morphological changes in the cells in the presence of 16:0, but not 18:2. Increasing concentrations of 16:0 (0.1-1.0 mM) caused gradual accumulation of intracellular membrane. Microvilli became strongly reduced in number. With 1.0 mM palmitic acid we found an increased incorporation of [1-14C]palmitic acid into phosphatidic acid (14.8% of total incorporation into phospholipid with 16:0 vs. < 0.5% with 18:2) and diacylglycerol (12.5% with 16:0 vs. < 0.5% with 18:2) and diacylglycerol (12.5% with 16:0 vs. 0.5% with 18:2) and the amount of intracellular phospholipid doubled. The morphological changes were completely reversed after 24 h with 1.0 mM 18:2. We conclude from our results that, compared to 18:2, 16:0 is not efficiently incorporated into triglycerides. 16:0 is incorporated into cellular phospholipids in a greater proportion than 18:2, causing accumulation of intracellular phospholipid and the precursors phosphatidic acid and diacylglycerol. Different processing of 18:2 and 16:0 by Caco-2 cells resulted in profound differences in triglyceride synthesis and lipoprotein composition and secretion.
利用肠道Caco - 2细胞的极化单层来研究饱和棕榈酸(16:0)和多不饱和亚油酸(18:2)对甘油三酯合成及脂蛋白分泌的影响。将单层细胞在生理浓度的棕榈酸(16:0)或亚油酸(18:2)存在下,于顶端或管腔侧孵育24小时。用1.0 mM的16:0或18:2孵育导致分泌的脂蛋白在组成和数量上存在差异。在与18:2孵育期间分泌的脂蛋白中的放射性标记脂质存在于乳糜微粒/极低密度脂蛋白(VLDL)密度范围内,而与16:0孵育时,分泌的脂蛋白处于中间密度/低密度脂蛋白(IDL/LDL)密度范围内。与1.0 mM的16:0(占总合成甘油三酯的18±3%)相比,在与1.0 mM的18:2孵育期间更多的甘油三酯分泌到(基底外侧)培养基中(占总合成甘油三酯的41±12%)。这些生化发现与16:0存在时细胞中明显的形态学变化相关,但18:2不存在这种情况。16:0浓度增加(0.1 - 1.0 mM)导致细胞内膜逐渐积累。微绒毛数量大幅减少。用1.0 mM棕榈酸时,我们发现[1 - 14C]棕榈酸掺入磷脂酸的比例增加(16:0时占总掺入磷脂的14.8%,而18:2时<0.5%)以及掺入二酰甘油的比例增加(16:0时为12..5%,18:2时<0.5%),并且细胞内磷脂的量增加了一倍。用1.0 mM的18:2处理24小时后,形态学变化完全逆转。我们从结果中得出结论,与18:2相比,16:0不能有效地掺入甘油三酯中。16:0比18:2更大比例地掺入细胞磷脂中,导致细胞内磷脂以及前体磷脂酸和二酰甘油的积累。Caco - 2细胞对18:2和16:0的不同处理导致甘油三酯合成、脂蛋白组成和分泌方面存在显著差异。