Guérin M, Dolphin P J, Chapman M J
INSERM Unit 321, Hôpital de la Pitié, Paris, France.
Arterioscler Thromb. 1994 Feb;14(2):199-206. doi: 10.1161/01.atv.14.2.199.
To date, several methods have been developed to determine the activity of plasma lipid transfer proteins. These methods have largely involved the addition of the transfer protein in question to labeled substrates, followed by prolonged incubation (4 to 18 hours) and subsequent evaluation of the radioactivity transferred to precipitated low-density lipoprotein (LDL). While adequate for determining the activity of cholesteryl ester transfer protein (CETP), these methods generally do not take into account the composition or levels of lipoproteins present within a given individual plasma because pools of high-density lipoprotein (HDL) are labeled and used for the transfer experiments. Both the direction and the extent of lipid transfer are dependent on the composition and relative abundance of both donor and acceptor particles as well as the activity of the lipid transfer protein(s). Here we describe a new method for the determination of the capacity of plasma samples to facilitate cholesteryl ester transfer from HDL to LDL and very-low-density lipoprotein (VLDL), a method that has several advantages. First, the subject's HDL is labeled and used for transfer. Second, the labeled HDL, in a quantity equivalent to 1% of the plasma HDL mass, is added to the subject's plasma, and therefore the relative abundance of both donor and acceptor particles is preserved at their physiological levels. Third, both cholesteryl ester mass and radioactivity are determined, allowing the net mass transfer of cholesteryl ester and cholesteryl ester exchange to be quantified separately.(ABSTRACT TRUNCATED AT 250 WORDS)
迄今为止,已经开发出几种方法来测定血浆脂质转运蛋白的活性。这些方法主要包括将相关转运蛋白添加到标记的底物中,然后长时间孵育(4至18小时),随后评估转移到沉淀的低密度脂蛋白(LDL)中的放射性。虽然这些方法足以测定胆固醇酯转运蛋白(CETP)的活性,但由于高密度脂蛋白(HDL)池被标记并用于转移实验,这些方法通常没有考虑给定个体血浆中脂蛋白的组成或水平。脂质转移的方向和程度取决于供体和受体颗粒的组成和相对丰度以及脂质转运蛋白的活性。在这里,我们描述了一种新的方法,用于测定血浆样品促进胆固醇酯从HDL转移到LDL和极低密度脂蛋白(VLDL)的能力,该方法具有几个优点。首先,将受试者的HDL进行标记并用于转移。其次,将相当于血浆HDL质量1%的标记HDL添加到受试者的血浆中,因此供体和受体颗粒的相对丰度保持在其生理水平。第三,同时测定胆固醇酯质量和放射性,从而可以分别量化胆固醇酯的净质量转移和胆固醇酯交换。(摘要截短于250字)