Schmid P C, Spimrova I, Schmid H H
Hormel Institute, University of Minnesota, Austin 55912, USA.
Lipids. 1997 Nov;32(11):1181-7. doi: 10.1007/s11745-997-0152-7.
Freshly isolated rat hepatocytes were incubated for 20 min with [U-14C]glycerol in the presence or absence of unlabeled linoleic (18:2n-6), arachidonic (20:4n-6), or docosahexaenoic (22:6n-3) acid, added as albumin complex in 10% ethanol. Most of the radioactivity (approximately 95%) recovered in hepatocyte lipids was present in phosphatidylcholine (PC), phosphatidylethanolamine (PE), and triacylglycerol (TAG). The presence of exogenous fatty acids resulted in (i) higher incorporation of [U-14C]glycerol, (ii) higher percentage of label in TAG, and (iii) enhanced formation of PC and PE molecular species bearing the exogenous fatty acid at both the sn-1 and sn-2 positions of glycerol. In each case, these molecular species contained 60 to 70% of the label in that lipid class. Further incubation of the cells for 40 and 80 min in the absence of labeled substrate and exogenous fatty acids resulted in a redistribution of label among PC and PE molecular species due to deacylation-reacylation at the sn-1 position of glycerol.
将新鲜分离的大鼠肝细胞与[U-14C]甘油在有无未标记的亚油酸(18:2n-6)、花生四烯酸(20:4n-6)或二十二碳六烯酸(22:6n-3)存在的情况下孵育20分钟,这些脂肪酸以白蛋白复合物的形式添加于10%乙醇中。肝细胞脂质中回收的大部分放射性(约95%)存在于磷脂酰胆碱(PC)、磷脂酰乙醇胺(PE)和三酰甘油(TAG)中。外源性脂肪酸的存在导致:(i)[U-14C]甘油的掺入增加;(ii)TAG中标记的百分比更高;(iii)在甘油的sn-1和sn-2位置带有外源性脂肪酸的PC和PE分子种类的形成增加。在每种情况下,这些分子种类在该脂质类别中含有60%至70%的标记。在没有标记底物和外源性脂肪酸的情况下,将细胞进一步孵育40分钟和80分钟,由于甘油sn-1位置的脱酰基-再酰基化作用,导致标记在PC和PE分子种类之间重新分布。