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CaCo-2细胞中的脂肪酸摄取与代谢:与微团或白蛋白结合存在的二十碳五烯酸(20:5(n-3))和油酸(18:1(n-9))

Fatty acid uptake and metabolism in CaCo-2 cells: eicosapentaenoic acid (20:5(n-3)) and oleic acid (18:1(n-9)) presented in association with micelles or albumin.

作者信息

Ranheim T, Gedde-Dahl A, Rustan A C, Drevon C A

机构信息

Section for Dietary Research, University of Oslo, Norway.

出版信息

Biochim Biophys Acta. 1994 Jun 2;1212(3):295-304. doi: 10.1016/0005-2760(94)90203-8.

Abstract

In vivo, long-chain fatty acids are incorporated into bile salt micelles, which solubilize the hydrophobic fatty acids before they are transported across the unstirred water layer to the intestinal brush border membrane. In the present study we have used CaCo-2 cells, cultured on filter membranes as a model of human enterocytes, and compared presentation of fatty acids bound to albumin with a micellar form. Absorption of eicosapentaenoic acid and oleic acid from micellar solutions was 4-times faster than from fatty acid-albumin complexes after 5 h incubation, and resulted in a corresponding increase in triacylglycerol synthesis and secretion. Mass determination of newly synthesized, cell-associated triacylglycerol after 5 h incubation, indicated a 5-fold increase in cells exposed to a micellar solution versus albumin-complexed fatty acids. A 2-fold larger fraction of the absorbed fatty acids was incorporated into triacylglycerol than into phospholipids when the fatty acids were presented as micelles. Analysis by resistive pulse technique showed that secreted lipoproteins of density less than 1.006 g/ml were in the same size-range as chylomicrons derived from human plasma. In spite of an increased amount of secreted triacylglycerol from cells supplemented with micellar fatty acids, there was no increase in the mean size of these particles. Synthesis and secretion of triacylglycerol in cells exposed to eicosapentaenoic acid and oleic acid were similar regardless of whether the fatty acids were presented to the cells associated with albumin or micelles, although the total amount of triacylglycerol synthesized and secreted was highest with micelles. When incubating CaCo-2 monolayers with eicosapentaenoic acid or oleic acid bound to albumin, a similar amount of radioactivity was released as CO2 and acid soluble products into the medium with less than 3% of the lipids being oxidized after 5 h of incubation. The oxidation rate of fatty acids in cells incubated with micelles was increased 40 to 100%. In conclusion, micellar fatty acids are absorbed, metabolized and influence secretion of lipoprotein particles to a higher extent than albumin-bound fatty acids in CaCo-2 cells, and there is no major difference between eicosapentaenoic acid and oleic acid.

摘要

在体内,长链脂肪酸会被整合到胆盐微胶粒中,这些微胶粒在疏水性脂肪酸穿过静止水层转运至肠刷状缘膜之前将其溶解。在本研究中,我们使用了培养在滤膜上的CaCo-2细胞作为人肠上皮细胞模型,并比较了与白蛋白结合的脂肪酸形式和微胶粒形式的脂肪酸呈现情况。在5小时孵育后,二十碳五烯酸和油酸从微胶粒溶液中的吸收速度比从脂肪酸-白蛋白复合物中的吸收速度快4倍,并导致三酰甘油合成和分泌相应增加。5小时孵育后对新合成的、与细胞相关的三酰甘油进行质量测定表明,与白蛋白复合脂肪酸相比,暴露于微胶粒溶液中的细胞中三酰甘油增加了5倍。当脂肪酸以微胶粒形式呈现时,被吸收的脂肪酸中整合到三酰甘油中的比例比整合到磷脂中的比例大2倍。电阻脉冲技术分析表明,密度小于1.006 g/ml的分泌脂蛋白与源自人血浆的乳糜微粒大小范围相同。尽管补充微胶粒脂肪酸的细胞分泌的三酰甘油量增加,但这些颗粒的平均大小没有增加。无论脂肪酸是以与白蛋白结合的形式还是微胶粒的形式呈现给细胞,暴露于二十碳五烯酸和油酸的细胞中三酰甘油的合成和分泌情况相似,尽管微胶粒形式下合成和分泌的三酰甘油总量最高。当用与白蛋白结合的二十碳五烯酸或油酸孵育CaCo-2单层细胞时,孵育5小时后,有相似量的放射性以二氧化碳和酸溶性产物的形式释放到培养基中,脂质氧化率不到3%。用微胶粒孵育的细胞中脂肪酸氧化率提高了40%至100%。总之,在CaCo-2细胞中,微胶粒脂肪酸比与白蛋白结合的脂肪酸在更高程度上被吸收、代谢并影响脂蛋白颗粒的分泌,并且二十碳五烯酸和油酸之间没有重大差异。

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