Gottlieb M H
Prep Biochem. 1978;8(6):463-9. doi: 10.1080/00327487808061663.
The in vitro extraction of cholesterol from erythrocytes by plasma lipoproteins of reduced cholesterol content would be expected to be free of cholesterol-unrelated alterations of the cell membrane. The earlier application of this method utilized whole blood plasma in which the major part of the lipoprotein cholesterol was esterified by the plasma enzyme lecithin-cholesterol acyl transferase (LCAT) in a preliminary incubation. Because of the cholesterol remaining unesterified in the plasma, only 35% of the cell cholesterol could be removed. The method reported here uses HDL3, a plasma lipoprotein which is the preferred substrate for LCAT, instead of whole plasma for the extraction. Multiple extractions with LCAT treated HDL3 resulted in the removal of up to 77% of the erythrocyte cholesterol with only minor hemolysis.
血浆脂蛋白从红细胞中体外提取胆固醇,若其胆固醇含量降低,则有望避免细胞膜发生与胆固醇无关的改变。该方法早期应用时使用的是全血血浆,其中大部分脂蛋白胆固醇在预孵育过程中被血浆酶卵磷脂胆固醇酰基转移酶(LCAT)酯化。由于血浆中仍有未酯化的胆固醇,所以只能去除35%的细胞胆固醇。此处报道的方法使用HDL3(一种血浆脂蛋白,是LCAT的首选底物)代替全血血浆进行提取。用经LCAT处理的HDL3进行多次提取,可去除高达77%的红细胞胆固醇,且仅有轻微溶血。