Smith Edward R
Department of Nephrology, The Royal Melbourne Hospital Kidney Care, 300 Grattan Street, Parkville, Melbourne, VIC, 3050, Australia.
Methods Mol Biol. 2016;1397:221-240. doi: 10.1007/978-1-4939-3353-2_15.
Multiple overlapping systemic and local inhibitory networks have evolved to prevent the unwanted deposition of mineral at ectopic sites. Fetuin-A is a liver-derived glycoprotein abundant in plasma that binds and stabilizes nascent mineral ion nuclei to form soluble colloidal high molecular weight complexes, called calciprotein particles (CPP). The binding of fetuin-A to mineral retards crystal ripening and precipitation from the aqueous phase, thereby facilitating the regulated clearance of mineral debris from the extracellular fluid. However, persistent disturbances in this humoral homeostatic system, as frequently seen in patients with Chronic Kidney Disease, may lead to the accumulation and aggregation of these nanoparticles in extraosseous tissues like the vasculature, driving inflammatory cascades, aberrant tissue remodeling, and functional impairment. Consistent with this conceptual framework, higher circulating CPP levels are associated with reduced renal function, increments in systemic inflammatory markers, derangements in bone morphogenetic cytokines, higher vascular calcification scores, aortic stiffening and an increased risk of death. This chapter describes optimized sample collection and preparative procedures for the isolation and enrichment of CPP from biological fluids. Methods for CPP quantitation are critically reviewed and detailed.
多种重叠的全身和局部抑制网络已经进化出来，以防止矿物质在异位部位的不必要沉积。胎球蛋白-A是一种源自肝脏的糖蛋白，在血浆中含量丰富，它能结合并稳定新生的矿质离子核，形成可溶性胶体高分子量复合物，称为钙蛋白颗粒（CPP）。胎球蛋白-A与矿物质的结合会延缓晶体成熟和从水相中沉淀，从而促进从细胞外液中对矿质碎片的调节性清除。然而，这种体液稳态系统的持续紊乱，如在慢性肾脏病患者中常见的那样，可能导致这些纳米颗粒在血管等骨外组织中积累和聚集，引发炎症级联反应、异常的组织重塑和功能损害。与这一概念框架一致，循环中较高的CPP水平与肾功能降低、全身炎症标志物增加、骨形态发生细胞因子紊乱、血管钙化评分升高、主动脉硬化以及死亡风险增加有关。本章描述了从生物流体中分离和富集CPP的优化样本采集和制备程序。对CPP定量方法进行了严格审查并详细说明。
