Tsuzuki Wakako
National Food Research Institute, Kannondai 2-1-12, Tsukuba, Ibaraki 305-8642, Japan.
Lipids. 2007 Jul;42(7):613-9. doi: 10.1007/s11745-007-3076-y. Epub 2007 Jun 21.
To elucidate the absorption characteristics of dietary lipids in the human intestine, we investigated the cellular uptake of lipid metabolites using a differential monolayer of the Caco2 cells. As lipid metabolites, several free fatty acids and 2-monoacylglycerols, were formed a mixed micelle by bile salts and lysophospholipids and they were supplied to the Caco2 cells. To estimate the effect of the mixed micelles on the permeability of cells' membranes during incubation with the mixed micelles, the transepitherial electrical resistance (TEER) value was monitored, and no pronounced changes of TEER was detected. This suggested that mixed micelles did not affect their cellular properties of the barrier measured by TEER. The lipid metabolites transferred from the mixed micelle into the Caco2 cells were determined quantitatively by an enzymatic colorimetric method and were done by thin layer chromatography (TLC) for a species of acylglycerols. These highly sensitive methods enabled us to monitor the transepithelial transports of various kinds of non-isotope-labeled various lipid metabolites. Newly re-synthesized triacylglycerols were accumulated in Caco2 cells after 30 min incubation with the mixed micelles, and their amounts increased gradually for 4 h. The secretion of re-esterified triacylglycerols into a basolateral medium from the Caco2 cells began at 2 h after the mixed micelles were added to the apical medium. The intake of external lipid metabolites by the Caco2 cells were evaluated by an initial 2-h incubation with the mixed micelles. For example, 2-monomyristin and 2-monopalmitin were more rapidly transferred into the Caco2 cells from the mixed micelles than 2-monocaprin was. On the other hand, the absorption rates of capric acid, lauric acid and myristic acid by the cells were larger than those of stearic acid and oleic acid. It revealed that the side-chain structure of these lipid metabolites affected their absorption by the Caco2 cells. The results of this study suggested that the Caco2 cell monolayer could be a useful model for investigating the involvement of dietary lipids in the transepithelial absorption in the human intestine.
为阐明人体肠道中膳食脂质的吸收特性,我们使用Caco2细胞的差异单层来研究脂质代谢物的细胞摄取。作为脂质代谢物,几种游离脂肪酸和2-单酰甘油与胆汁盐和溶血磷脂形成混合微团,并将它们提供给Caco2细胞。为评估混合微团在与混合微团孵育期间对细胞膜通透性的影响,监测跨上皮电阻(TEER)值,未检测到TEER有明显变化。这表明混合微团不影响通过TEER测量的其屏障的细胞特性。从混合微团转移到Caco2细胞中的脂质代谢物通过酶比色法进行定量测定,对于一种酰基甘油则通过薄层色谱法(TLC)进行测定。这些高灵敏度方法使我们能够监测各种非同位素标记的各种脂质代谢物的跨上皮转运。与混合微团孵育30分钟后,新重新合成的三酰甘油在Caco2细胞中积累,并且其含量在4小时内逐渐增加。从混合微团添加到顶侧培养基开始2小时后,重新酯化的三酰甘油从Caco2细胞分泌到基底外侧培养基中。通过与混合微团最初孵育2小时来评估Caco2细胞对外部脂质代谢物的摄取。例如,2-肉豆蔻酸甘油酯和2-棕榈酸甘油酯比2-癸酸甘油酯更快地从混合微团转移到Caco2细胞中。另一方面,细胞对癸酸、月桂酸和肉豆蔻酸的吸收率大于硬脂酸和油酸。结果表明,这些脂质代谢物的侧链结构影响它们被Caco2细胞的吸收。本研究结果表明,Caco2细胞单层可能是研究膳食脂质在人体肠道跨上皮吸收中作用的有用模型。