Yu F G, Ando S, Hayashi S
Laboratory of Food Chemistry, Faculty of Fisheries, Kagoshima University, Japan.
Cell Struct Funct. 1991 Aug;16(4):347-55. doi: 10.1247/csf.16.347.
The lipoprotein secreted by cultured eel hepatocytes was fractionated by density gradient ultracentrifugation and compared with eel serum lipoproteins. Eel hepatocytes were cultured for 7 to 10 days as a monolayer in Williams' medium E containing 5% fetal bovine serum and 0.16 microM insulin on a dish precoated with fibronectin of horse serum. The only lipoprotein secreted by eel hepatocytes was a very-low-density lipoprotein like one which consisted of 69% triglyceride, 15% phospholipid, 4% cholesterol, and 12% protein. On the other hand, very-low-density lipoprotein and high density lipoprotein were found in eel serum, in which high density lipoprotein was a main lipoprotein. The secreted lipoprotein contained apo B and apo A as the main protein components. Furthermore, the lipoprotein contained proapo A-I in addition to apo A-I, which was proved by comparing the amino acid composition of both proteins. In our discussion, we noted that the lipoprotein secreted by eel hepatocytes was a good material for the study of high-density lipoprotein formation.
通过密度梯度超速离心法对培养的鳗鱼肝细胞分泌的脂蛋白进行分级分离,并与鳗鱼血清脂蛋白进行比较。将鳗鱼肝细胞在预先涂有马血清纤连蛋白的培养皿中,于含有5%胎牛血清和0.16微摩尔胰岛素的威廉姆斯E培养基中单层培养7至10天。鳗鱼肝细胞分泌的唯一脂蛋白是一种极低密度脂蛋白样物质,其由69%的甘油三酯、15%的磷脂、4%的胆固醇和12%的蛋白质组成。另一方面,在鳗鱼血清中发现了极低密度脂蛋白和高密度脂蛋白,其中高密度脂蛋白是主要的脂蛋白。分泌的脂蛋白以载脂蛋白B和载脂蛋白A作为主要蛋白质成分。此外,通过比较两种蛋白质的氨基酸组成证明,该脂蛋白除了载脂蛋白A-I外还含有前载脂蛋白A-I。在我们的讨论中,我们指出鳗鱼肝细胞分泌的脂蛋白是研究高密度脂蛋白形成的良好材料。