Noël S P, Dupras R
Biochim Biophys Acta. 1983 Nov 29;754(2):117-25. doi: 10.1016/0005-2760(83)90152-2.
The aim of this study was to determine the kinetic parameters of the hepatic uptake of VLDL remnant cholesteryl esters. Rat livers were perfused in situ with a broad range of remnant [3H]cholesteryl ester concentrations of known specific radioactivity. Following exactly 3 min of perfusion, hepatic lipids were extracted and labelled cholesteryl esters were separated by thin-layer chromatography and counted. The rate of cholesteryl ester uptake was a saturable process and the apparent kinetic parameters were determined from the Lineweaver-Burk plot of the data. Km and Vmax were calculated to be 72 microM and 35 nmol cholesteryl ester/min per g liver, respectively. For the purpose of comparison, we have expressed our kinetic parameters in terms of number of particles (Vmax = 0.022 nmol particles/min per g liver and Km = 45 nM) and compared our values with those obtained with chylomicron remnants by another group of investigators (Sherrill, B.C., Innerarity, T.L. and Mahley, R.W. (1980) J. Biol. Chem. 255, 1804-1807). We found that the maximal capacity for the removal of VLDL particles was similar to what was observed with rat chylomicron remnants. In contrast, the Km for the uptake process of VLDL remnant particles was approximately four times higher than that of rat chylomicron remnant particles. Our results are consistent with the hypothesis that hepatic removal of both chylomicron and VLDL remnants is mediated by the same receptor, but suggest that the affinity of VLDL remnants for the hepatic removal process is substantially lower, possibly due to structural differences between the two remnant particles.
本研究的目的是确定极低密度脂蛋白(VLDL)残余胆固醇酯肝摄取的动力学参数。用一系列已知比放射性的残余[³H]胆固醇酯浓度对大鼠肝脏进行原位灌注。灌注恰好3分钟后,提取肝脏脂质,通过薄层层析分离标记的胆固醇酯并进行计数。胆固醇酯摄取速率是一个可饱和过程,根据数据的Lineweaver-Burk图确定表观动力学参数。计算得出Km和Vmax分别为72微摩尔和每克肝脏每分钟35纳摩尔胆固醇酯。为了进行比较,我们以颗粒数量表示动力学参数(Vmax = 每克肝脏每分钟0.022纳摩尔颗粒,Km = 45纳摩尔),并将我们的值与另一组研究人员用乳糜微粒残余物获得的值进行比较(谢里尔,B.C.,英纳里蒂,T.L.和马利,R.W.(1980年)《生物化学杂志》255,1804 - 1807)。我们发现去除VLDL颗粒的最大能力与在大鼠乳糜微粒残余物中观察到的相似。相比之下,VLDL残余颗粒摄取过程的Km大约是大鼠乳糜微粒残余颗粒的四倍。我们的结果与以下假设一致,即乳糜微粒和VLDL残余物的肝脏清除均由同一受体介导,但表明VLDL残余物对肝脏清除过程的亲和力显著较低,这可能是由于两种残余颗粒之间的结构差异所致。