Abstract：
Lipoproteins in plasma are constituted by the least dense chylomicron, very-low-density lipoprotein (VLDL), low-density lipoprotein (LDL), and high-density lipoprotein (HDL) that can be separated using commercially available medium such as iodixanol. Iodixanol constitutes the self-generated density gradient to fractionate lipoproteins by rapid ultracentrifugation method, replacing time-consuming protocols. Filling the centrifuge tubes is technically easier and faster than layering salt gradients and is reproducible. The separated lipoproteins by this method are closest to the native state with 80 to 100% recovery possible. Low-density lipoprotein is the major carrier of cholesterol in systemic circulation. The plasma isolated LDL is purified to be used as native LDL and for the preparation of oxidized LDL (oxLDL). The oxLDL is characterized for its oxidation, by various methods based on assay of the lipid and protein oxidation products such as TBARS, conjugated diene formation, and by other methods such as agarose gel electrophoresis. Rapid isolation of LDL particles from human plasma is useful for lipid peroxidation studies, characterization of subclass for functional studies and clinical correlation especially in cardiovascular diseases apart from lipidomic, and proteomic studies. OxLDL preparations are done in vitro chiefly based on copper-induced oxidation; glucose and other prooxidants. Which are used for various studies using animal model and in vitro cell models especially to understand macrophage-mediated atheroma formation, vascular endothelial cell dysfunction, cell signaling studies has scope for extensive research in metabolic dysfunction of various cells. This chapter deals with one of the applications in the in vitro cell models using macrophage (THP-1 cell line) and human retinal pigment epithelial cell (ARPE-19 cell line) to study the oxLDL uptake using fluorescently labeled oxidized LDL (DiI-oxLDL).
摘要：
血浆中的脂蛋白由密度最小的乳糜微粒构成，极低密度脂蛋白（VLDL），低密度脂蛋白（LDL），和高密度脂蛋白（HDL），可以使用市售培养基如碘克沙醇分离。碘克沙醇构成自生成密度梯度，通过快速超速离心方法分离脂蛋白，替换耗时的协议。填充离心管在技术上比分层盐梯度更容易和更快,并且是可再现的。通过该方法分离的脂蛋白最接近天然状态,可能有80-100%的回收率。低密度脂蛋白是全身循环中胆固醇的主要载体。纯化血浆分离的LDL以用作天然LDL并用于制备氧化LDL(oxLDL)。oxLDL的特征在于其氧化，通过基于脂质和蛋白质氧化产物如TBARS测定的各种方法，共轭二烯的形成，和其他方法，如琼脂糖凝胶电泳。从人血浆中快速分离LDL颗粒可用于脂质过氧化研究，功能研究和临床相关性的亚类表征，特别是在除了脂质组学之外的心血管疾病中，和蛋白质组学研究。OxLDL制剂主要基于铜诱导的氧化、葡萄糖和其他促氧化剂在体外进行。使用动物模型和体外细胞模型进行各种研究，特别是了解巨噬细胞介导的动脉粥样硬化形成，血管内皮细胞功能障碍，细胞信号研究具有广泛研究各种细胞代谢功能障碍的空间。本章涉及使用巨噬细胞（THP-1细胞系）和人视网膜色素上皮细胞（ARPE-19细胞系）在体外细胞模型中的应用之一，以研究使用荧光标记的氧化LDL（DiI-oxLDL）的oxLDL摄取。
