Li Kang, Wong David K, Luk Fu Sang, Kim Roy Y, Raffai Robert L
Department of Surgery, University of California & VA Medical Center, San Francisco, CA, USA.
Methods Mol Biol. 2018;1740:139-153. doi: 10.1007/978-1-4939-7652-2_11.
Plasma lipoproteins are essential vehicles of lipid distribution for cellular energy and structural requirements as well as for excretion of lipid excess. Imbalances in lipoprotein metabolism are known to contribute to metabolic diseases ranging from vascular inflammation and atherosclerosis to obesity and diabetes. The lipid and protein cargo carried by lipoprotein subclasses have long been the focus of studies exploring the contribution of plasma lipoproteins in health and in metabolic disorders. More recent studies have revealed the presence of noncoding RNA as a new form of cargo carried by plasma lipoproteins. Lipoprotein-associated microRNAs have been identified to distribute differentially among plasma lipoprotein subclasses and contribute to cellular signaling. These findings highlight plasma lipoprotein-associated RNA as a potential source of biological signaling and warrant a renewed interest in the study of plasma lipoprotein biology. This chapter describes principles and methods based on density ultracentrifugation and size exclusion chromatography for the isolation of plasma lipoproteins as a source of extracellular RNA.
血浆脂蛋白是脂质分布的重要载体,用于满足细胞能量和结构需求以及排出多余脂质。已知脂蛋白代谢失衡会导致从血管炎症、动脉粥样硬化到肥胖症和糖尿病等一系列代谢性疾病。脂蛋白亚类所携带的脂质和蛋白质成分长期以来一直是探索血浆脂蛋白在健康和代谢紊乱中作用的研究重点。最近的研究发现,非编码RNA是血浆脂蛋白携带的一种新成分。已鉴定出脂蛋白相关的微小RNA在血浆脂蛋白亚类之间存在差异分布,并参与细胞信号传导。这些发现突出了血浆脂蛋白相关RNA作为生物信号潜在来源的重要性,值得重新关注血浆脂蛋白生物学的研究。本章介绍基于密度超速离心和尺寸排阻色谱法的原理和方法,用于分离血浆脂蛋白以获取细胞外RNA。