Cross K E, Dodds P F
Department of Biochemistry and Biological Sciences, Wye College (University of London), Kent, U.K.
Biochem J. 1988 Oct 1;255(1):259-65.
(1) A method has been developed to separate hepatocytes, isolated from laying hens, according to their densities, using discontinuous density-gradient centrifugation on Nycodenz. (2) The hepatocytes recovered from the interface of the 5% and 10% Nycodenz layers were rich in triacylglycerol and were termed 'fatty' hepatocytes: 'non-fatty' hepatocytes were obtained from the interface of the 15% and 30% Nycodenz layers and contained less than one-quarter as much triacylglycerol. (3) 'Fatty' hepatocytes incorporated radiolabelled glucose and glycerol into total lipid at more than twice the rate of 'non-fatty' cells: the corresponding increases in the incorporation of radiolabelled choline and valine into phospholipid and protein respectively were smaller and not statistically significant. (4) A higher proportion of glycerol and glucose incorporated into total lipid was found to be phospholipid in the 'non-fatty' hepatocytes. (5) A higher proportion of radiolabelled lipid or protein formed from glycerol or valine respectively was secreted into the medium by the 'non-fatty' hepatocytes. (6) The use of these hepatocytes as a model to study fatty liver syndromes is discussed.
(1) 已开发出一种方法,通过在 Nycodenz 上进行不连续密度梯度离心,根据密度分离从产蛋母鸡中分离出的肝细胞。(2) 从 5% 和 10% Nycodenz 层界面回收的肝细胞富含三酰甘油,被称为“脂肪性”肝细胞:“非脂肪性”肝细胞从 15% 和 30% Nycodenz 层界面获得,其所含三酰甘油不到前者的四分之一。(3) “脂肪性”肝细胞将放射性标记的葡萄糖和甘油掺入总脂质的速率是“非脂肪性”细胞的两倍多:放射性标记的胆碱和缬氨酸分别掺入磷脂和蛋白质的相应增加较小且无统计学意义。(4) 在“非脂肪性”肝细胞中,掺入总脂质的甘油和葡萄糖中较高比例是磷脂。(5) “非脂肪性”肝细胞分别由甘油或缬氨酸形成的较高比例的放射性标记脂质或蛋白质分泌到培养基中。(6) 讨论了使用这些肝细胞作为研究脂肪肝综合征模型的情况。