Kilsdonk E P, Yancey P G, Stoudt G W, Bangerter F W, Johnson W J, Phillips M C, Rothblat G H
Department of Biochemistry, Medical College of Pennsylvania, Hahnemann University, Philadelphia 19129, USA.
J Biol Chem. 1995 Jul 21;270(29):17250-6. doi: 10.1074/jbc.270.29.17250.
In this study, we compared cholesterol efflux mediated by either high density lipoproteins (HDL3) or beta-cyclodextrins, cyclic oligosaccharides that are able to dissolve lipids in their hydrophobic core. beta-Cyclodextrin, 2-hydroxypropyl-beta-cyclodextrin, and methyl-beta-cyclodextrin at 10 mM induced the release of 50-90% of L-cell [3H]cholesterol after 8 h of incubation, with a major portion of this cholesterol being released in the first 1-2 h of incubation. The cholesterol efflux kinetics are different if cells are incubated with HDL3, which induces a relatively constant rate of release of cholesterol throughout an 8-h incubation. Cholesterol efflux to cyclodextrins was much greater than phospholipid release. To test the hypothesis that maximal efflux rate constants for a particular cell are independent of the type of acceptor, we estimated the maximal rate constants for efflux (Vmax) of cellular cholesterol from L-cells, Fu5AH cells, and GM3468A fibroblasts. The rate constant for HDL3-mediated efflux varied among cell lines in the order Fu5AH > L-cells > fibroblasts. However, these differences were not evident when cyclodextrins were used as cholesterol acceptors. The estimated Vmax values for cyclodextrin-mediated efflux were 3.5-70-fold greater than for HDL3 for the three cell lines. The very high efficiency of cyclodextrins in stimulating cell cholesterol efflux suggests that these compounds can be used in two general ways for studies of atherosclerosis: 1) as research tools to probe mechanisms of cholesterol transport and aspects of membrane structure or 2) as potential pharmacological agents that could modify in vivo cholesterol metabolism and influence the development of the atherosclerotic plaque.
在本研究中,我们比较了由高密度脂蛋白(HDL3)或β-环糊精介导的胆固醇流出,β-环糊精是一种能够在其疏水核心中溶解脂质的环状寡糖。10 mM的β-环糊精、2-羟丙基-β-环糊精和甲基-β-环糊精在孵育8小时后诱导L细胞[3H]胆固醇释放50-90%,其中大部分胆固醇在孵育的前1-2小时内释放。如果细胞与HDL3孵育,胆固醇流出动力学则不同,HDL3在整个8小时的孵育过程中诱导胆固醇以相对恒定的速率释放。胆固醇向环糊精的流出远大于磷脂的释放。为了检验特定细胞的最大流出速率常数与受体类型无关这一假设,我们估计了L细胞、Fu5AH细胞和GM3468A成纤维细胞中细胞胆固醇流出的最大速率常数(Vmax)。HDL3介导的流出速率常数在不同细胞系中的顺序为Fu5AH>L细胞>成纤维细胞。然而,当使用环糊精作为胆固醇受体时,这些差异并不明显。对于这三种细胞系,环糊精介导的流出的估计Vmax值比HDL3介导的流出大3.5-70倍。环糊精在刺激细胞胆固醇流出方面的极高效率表明,这些化合物可用于动脉粥样硬化研究的两种一般方式:1)作为研究胆固醇转运机制和膜结构方面的研究工具;2)作为可能改变体内胆固醇代谢并影响动脉粥样硬化斑块发展的潜在药物制剂。