Preobrazhensky S, Trakht I, Chestkov V, Wentz M
Gull Laboratories, Inc., Salt Lake City, Utah 84177, USA.
Anal Biochem. 1995 May 1;227(1):225-34. doi: 10.1006/abio.1995.1274.
Numerous reports indicate that the oxidation of low-density lipoprotein (LDL) can significantly change its metabolic and physiological properties. Most methods for evaluation of LDL oxidation require isolation of lipoprotein, making the procedure laborious and increasing the probability of artifactual modification of LDL. In this paper we describe an immunochemical approach which can be used to measure the oxidation of isolated LDL and apoprotein B in unfractionated serum and to evaluate the effects of antioxidants on these processes. The procedure is based on differential recognition by monoclonal antibodies of native and oxidized lipoproteins. The results obtained with our assay indicate a strong correlation between the changes of apo B epitope expression during oxidation and the formation of conjugated dienes, changes in lipoprotein electrophoretic mobility, and interaction with fibroblast and macrophage receptors. The sensitivity of apo B to oxidation varies greatly among serum samples obtained from individual donors. These differences do not correlate with the differences in sensitivity to oxidation of LDL isolated from the blood samples of the same donors. It is also shown that apo B oxidation in serum can be progressively inhibited in the presence of increasing amounts of various antioxidants.
大量报告表明，低密度脂蛋白（LDL）的氧化可显著改变其代谢和生理特性。大多数评估LDL氧化的方法都需要分离脂蛋白，这使得该过程繁琐，并增加了LDL人为修饰的可能性。在本文中，我们描述了一种免疫化学方法，该方法可用于测量未分级血清中分离的LDL和载脂蛋白B的氧化，并评估抗氧化剂对这些过程的影响。该方法基于单克隆抗体对天然脂蛋白和氧化脂蛋白的差异识别。我们的检测结果表明，氧化过程中载脂蛋白B表位表达的变化与共轭二烯的形成、脂蛋白电泳迁移率的变化以及与成纤维细胞和巨噬细胞受体的相互作用之间存在很强的相关性。从个体供体获得的血清样本中，载脂蛋白B对氧化的敏感性差异很大。这些差异与从相同供体血液样本中分离的LDL对氧化的敏感性差异无关。研究还表明，在存在越来越多的各种抗氧化剂的情况下，血清中载脂蛋白B的氧化可被逐步抑制。
