Oliveira H C, Quintão E C
Lipids Lab. University of São Paulo Medical School, Brazil.
J Biochem Biophys Methods. 1996 Apr;32(1):45-57. doi: 10.1016/0165-022x(94)00060-q.
The cholesteryl ester flow between high-density lipoproteins (HDL) and triglyceride-rich lipoproteins was investigated utilizing HDL fractions and synthetic lipid emulsion particles (EM). HDL was labeled in vitro with [1,2-3H(n)]cholesteryl ester or with [1,2,6,7-3H(n)]cholesteryl oleate, whereas EM were made with [4-14C]cholesteryl oleate (CO) or [carboxyl-14C]triolein (TO). The cholesteryl ester (CE) transfer rate between HDL and EM was spontaneous to some extent (because it occurred in saline medium), saturable, enhanced in a dose-dependent manner by the plasma fraction at D > 1.21 g/ml ascribed to its CETP activity, and greater for HDL3 than for HDL2. Unesterified fatty acids in the medium elicited two opposing effects: (1) enhanced the spontaneous cholesteryl oleate shift to EM and inhibited the reverse flow from EM to HDL3 but not to HDL2; (2) partially impaired the plasma D > 1.21 g/ml-induced bidirectional cholesteryl oleate flow. Approx. 2 mol TO from EM exchange for 1 mol CO from HDL3. Net cholesteryl ester balance was dependent on the concentration of HDL-cholesteryl ester, and independent from EM-cholesteryl oleate, possibly due to the much smaller concentration of the latter. These in vitro experiments shed light on the complex physical chemistry of transport of cholesteryl ester and triolein between HDL and TG-rich lipoproteins which occurs during the metabolism of chylomicrons in plasma.
利用高密度脂蛋白(HDL)组分和合成脂质乳剂颗粒(EM)研究了高密度脂蛋白与富含甘油三酯的脂蛋白之间的胆固醇酯流动情况。HDL在体外用[1,2-³H(n)]胆固醇酯或[1,2,6,7-³H(n)]油酸胆固醇酯进行标记,而EM则用[4-¹⁴C]油酸胆固醇酯(CO)或[羧基-¹⁴C]三油精(TO)制备。HDL与EM之间的胆固醇酯(CE)转移率在一定程度上是自发的(因为它发生在盐溶液介质中),具有饱和性,在D>1.21 g/ml的血浆组分因其胆固醇酯转移蛋白(CETP)活性而以剂量依赖性方式增强,且HDL3的转移率高于HDL2。培养基中的未酯化脂肪酸产生两种相反的作用:(1)增强油酸胆固醇酯向EM的自发转移,并抑制从EM到HDL3但不抑制到HDL2的逆向流动;(2)部分损害血浆D>1.21 g/ml诱导的双向油酸胆固醇酯流动。约2摩尔来自EM的TO与1摩尔来自HDL3的CO进行交换。胆固醇酯的净平衡取决于HDL-胆固醇酯的浓度,且与EM-油酸胆固醇酯无关,这可能是由于后者的浓度要小得多。这些体外实验揭示了在血浆中乳糜微粒代谢过程中HDL与富含甘油三酯的脂蛋白之间胆固醇酯和三油精转运的复杂物理化学过程。