Alpers D H, Lock D R, Lancaster N, Poksay K, Schonfeld G
J Lipid Res. 1985 Jan;26(1):1-10.
Chylomicrons and very low density lipoproteins (VLDL) are produced by the intestine and these nascent particles are thought to be similar to their counterparts in intestinal lymph. To study the relationship between these lipoproteins within the cell and those secreted into the lamina propria and lymph, we have isolated enterocytes, lamina propria, and mesenteric lymph from rats while fasted and after corn oil feeding. Apolipoprotein A-I and B content were measured by radioimmunoassay in cell, lamina propria, and lymph fractions separated by Sepharose 6B and 10% agarose chromatography, and by KBr isopycnic density centrifugation. ApoA-I in the cell and the underlying lamina propria was found partly in those fractions in which chylomicron and very low density lipoproteins (chylo-VLDL) and high density lipoproteins (HDL) elute, but more abundantly where unassociated 125I-labeled apoA-I was eluted. In the lymph, however, 74% of apoA-I eluted in the HDL region and no peak of free apoA-I was found. ApoB and apoC-III within the enterocyte were found distributed in the position of particles eluting not only with chylomicrons and VLDL, but also in the regions corresponding to LDL and HDL. In the lamina propria and lymph, on the other hand, most of the apoB was found in the region of VLDL and chylomicrons. These results indicate that the patterns in lymph lipoproteins and the lamina propria do not exactly mirror the distribution of apoA-I and B among lipoproteins inside the cell. This may be because intracellular apoproteins may be unassociated with lipoproteins, or they could be associated with lipoproteins in various stages of assembly of protein with lipids. Furthermore, the apoprotein composition of intestinal lipoproteins is altered after secretion from the enterocyte. Finally, not all apoproteins seem to be secreted in association with identifiable lipoprotein particles from the enterocyte.
乳糜微粒和极低密度脂蛋白(VLDL)由肠道产生,这些新生颗粒被认为与肠淋巴中的对应物相似。为了研究细胞内这些脂蛋白与分泌到固有层和淋巴中的脂蛋白之间的关系,我们在大鼠禁食和喂食玉米油后分离了肠上皮细胞、固有层和肠系膜淋巴。通过放射免疫分析法测定了经琼脂糖6B和10%琼脂糖色谱法以及KBr等密度离心法分离得到的细胞、固有层和淋巴组分中的载脂蛋白A-I和B含量。发现细胞和下层固有层中的载脂蛋白A-I部分存在于乳糜微粒、极低密度脂蛋白(乳糜微粒 - VLDL)和高密度脂蛋白(HDL)洗脱的组分中,但在未结合的125I标记载脂蛋白A-I洗脱的位置更为丰富。然而,在淋巴中,74%的载脂蛋白A-I在HDL区域洗脱,未发现游离载脂蛋白A-I的峰。发现肠上皮细胞内的载脂蛋白B和载脂蛋白C-III不仅分布在与乳糜微粒和VLDL一起洗脱的颗粒位置,还分布在与低密度脂蛋白(LDL)和HDL相对应的区域。另一方面,在固有层和淋巴中,大部分载脂蛋白B存在于VLDL和乳糜微粒区域。这些结果表明,淋巴脂蛋白和固有层中的模式并不完全反映细胞内载脂蛋白A-I和B在脂蛋白之间的分布。这可能是因为细胞内载脂蛋白可能未与脂蛋白结合,或者它们可能与蛋白质与脂质组装的各个阶段的脂蛋白结合。此外,肠脂蛋白的载脂蛋白组成在从肠上皮细胞分泌后会发生改变。最后,并非所有载脂蛋白似乎都是与从肠上皮细胞中可识别的脂蛋白颗粒一起分泌的。