Fluiter K, Vietsch H, Biessen E A, Kostner G M, van Berkel T J, Sattler W
Division of Biopharmaceutics, Leiden/Amsterdam Center for Drug Research, University of Leiden, Sylvius Laboratories, The Netherlands.
Biochem J. 1996 Oct 15;319 ( Pt 2)(Pt 2):471-6. doi: 10.1042/bj3190471.
Oxidation of low-density lipoprotein (LDL) leads initially to the formation of LDL-associated cholesteryl ester hydroperoxides (CEOOH). LDL-associated CEOOH can be transferred to high-density lipoprotein (HDL), and HDL-associated CEOOH are rapidly reduced to the corresponding hydroxides (CEOH) by an intrinsic peroxidase-like activity. We have now performed in vivo experiments to quantify the clearance rates and to identify the uptake sites of HDL-associated [3H]Ch18:2-OH in rats. Upon injection into rats, HDL-associated [3H]Ch18:2-OH is removed more rapidly from the circulation than HDL-associated [3H]Ch18:2. Two minutes after administration of [3H]Ch18:2-OH-HDL, 19.6 +/- 2.6% (S.E.M.; n = 4) of the label was taken up by the liver as compared with 2.4 +/- 0.25% (S.E.M.; n = 4) for [3H]Ch18:2-HDL. Organ distribution studies indicated that only the liver and adrenals exhibited preferential uptake of [3H]Ch18:2-OH as compared with [3H]Ch18:2, with the liver as the major site of uptake. A cell-separation procedure, employed 10 min after injection of [3H]Ch18:2-OH-HDL or [3H]Ch18:2-HDL, demonstrated that within the liver only parenchymal cells take up HDL-CE by the selective uptake pathway. Selective uptake by parenchymal cells of [3H]Ch18:2-OH was 3-fold higher than that of [3H]Ch18:2, while Kupffer and endothelial cell uptake of the lipid tracers reflected HDL holoparticle uptake (as analysed with iodinated versus cholesteryl ester-labelled HDL). The efficient uptake of [3H]Ch18:2-OH by parenchymal cells was coupled to a 3-fold increase in rate of radioactive bile acid secretion from [3H]Ch18:2-OH-HDL as compared with [3H]Ch18:2-HDL. In vitro studies with freshly isolated parenchymal cells showed that the association of [3H]Ch18:2-OH-HDL at 37 degrees C exceeded [3H]Ch18:2-HDL uptake almost 4-fold. Our results indicate that HDL-associated CEOH are efficiently and selectively removed from the blood circulation by the liver in vivo. The selective liver uptake is specifically exerted by parenchymal cells and coupled to a rapid biliary secretion pathway. The liver uptake and biliary secretion route may allow HDL to function as an efficient protection system against potentially atherogenic CEOOH.
低密度脂蛋白(LDL)的氧化最初会导致形成与LDL相关的胆固醇酯氢过氧化物(CEOOH)。与LDL相关的CEOOH可转移至高密度脂蛋白(HDL),而与HDL相关的CEOOH会通过一种内在的类过氧化物酶活性迅速还原为相应的氢氧化物(CEOH)。我们现已进行体内实验,以量化HDL相关的[3H]Ch18:2 - OH在大鼠体内的清除率,并确定其摄取部位。将HDL相关的[3H]Ch18:2 - OH注射入大鼠后,其从循环中清除的速度比HDL相关的[3H]Ch18:2更快。给予[3H]Ch18:2 - OH - HDL两分钟后,肝脏摄取了19.6±2.6%(标准误;n = 4)的标记物,而[3H]Ch18:2 - HDL的摄取率为2.4±0.25%(标准误;n = 4)。器官分布研究表明,与[3H]Ch18:2相比,只有肝脏和肾上腺对[3H]Ch18:2 - OH有优先摄取,其中肝脏是主要摄取部位。在注射[3H]Ch18:2 - OH - HDL或[3H]Ch18:2 - HDL 10分钟后采用的细胞分离程序表明,在肝脏内只有实质细胞通过选择性摄取途径摄取HDL - CE。实质细胞对[3H]Ch18:2 - OH的选择性摄取比[3H]Ch18:2高3倍,而库普弗细胞和内皮细胞对脂质示踪剂的摄取反映了HDL全颗粒的摄取(用碘化与胆固醇酯标记的HDL进行分析)。与[3H]Ch18:2 - HDL相比,实质细胞对[3H]Ch18:2 - OH的有效摄取与放射性胆汁酸从[3H]Ch18:2 - OH - HDL分泌的速率增加3倍相关。对新鲜分离的实质细胞进行的体外研究表明,在37℃时[3H]Ch18:2 - OH - HDL的结合量几乎比[3H]Ch18:2 - HDL的摄取量高4倍。我们的结果表明,在体内,肝脏能有效地、选择性地从血液循环中清除与HDL相关的CEOH。肝脏的选择性摄取是由实质细胞特异性发挥作用的,并与快速的胆汁分泌途径相关。肝脏摄取和胆汁分泌途径可能使HDL发挥有效的保护系统作用,抵御潜在的致动脉粥样硬化的CEOOH。