Preobrazhensky S N, Tsibulsky V P, Fuki I V, Ivanov V O, Repin V S, Smirnov V N
Anal Biochem. 1986 May 1;154(2):382-7. doi: 10.1016/0003-2697(86)90002-3.
Mouse macrophages (line J 774) were incubated with monospecific goat anti-low density lipoprotein antibodies, which were conjugated to horseradish peroxidase (AB-HRP). Addition of low density lipoprotein (LDL) modified by treatment with malondialdehyde to cultures of these cells resulted in a dose-dependent increase in the amount of cell-associated enzyme activity. The concentration curve was hyperbolic with half-saturation of modified LDL at a concentration of about 3 micrograms/ml. This effect was completely blocked by polyinosinic acid and was not observed in experiments with human fibroblasts, which do not exhibit high affinity binding sites that recognize chemically modified LDL. Our data indicate that receptor-mediated endocytosis of AB-HRP in the presence of native or modified LDL may be used as very simple, efficient, and sensitive assay for investigation of the scavenger receptors for modified LDL.
将小鼠巨噬细胞（J 774系）与与辣根过氧化物酶偶联的单特异性山羊抗低密度脂蛋白抗体（AB-HRP）一起孵育。向这些细胞培养物中添加经丙二醛处理修饰的低密度脂蛋白（LDL），导致细胞相关酶活性量呈剂量依赖性增加。浓度曲线呈双曲线，修饰的LDL在约3微克/毫升浓度时达到半饱和。这种效应被聚肌苷酸完全阻断，并且在用人成纤维细胞进行的实验中未观察到，人成纤维细胞不表现出识别化学修饰LDL的高亲和力结合位点。我们的数据表明，在天然或修饰的LDL存在下，AB-HRP的受体介导的内吞作用可作为一种非常简单、有效且灵敏的检测方法，用于研究修饰LDL的清道夫受体。
