Escolà-Gil Joan Carles, Lee-Rueckert Miriam, Santos David, Cedó Lídia, Blanco-Vaca Francisco, Julve Josep
Institut de Recerca de l'Hospital de la Santa Creu i Sant Pau, Institut d'Investigacions Biomèdiques (IIB) Sant Pau, C/Antoni M. Claret 167, 08025, Barcelona, Spain.
CIBER de Diabetes y Enfermedades Metabólicas Asociadas. CIBERDEM, Madrid, Spain.
Methods Mol Biol. 2015;1339:211-33. doi: 10.1007/978-1-4939-2929-0_15.
Promotion of reverse cholesterol transport (RCT) is thought to be a major HDL-mediated mechanism for protecting against atherosclerosis. Preclinical studies support the concept that increasing cholesterol efflux from macrophages may confer atheroprotective benefits independently of the plasma HDL-cholesterol concentration. The application of the macrophage-to-feces RCT method in genetically engineered mice has provided evidence that this major HDL property correlates closely with changes in atherosclerosis susceptibility. This chapter provides details on the methodologies currently used to measure in vitro cholesterol efflux from macrophages or in vivo macrophage-specific RCT. The general principles and techniques described herein may be applied to measure the in vitro cholesterol efflux capacity of human serum in macrophage cultures and to evaluate the effect of different experimental pathophysiological conditions or the efficacy of different therapeutic strategies on the modulation of in vivo macrophage-RCT in mice.
促进胆固醇逆向转运(RCT)被认为是高密度脂蛋白(HDL)介导的预防动脉粥样硬化的主要机制。临床前研究支持这样一种观点,即增加巨噬细胞的胆固醇流出可能独立于血浆HDL胆固醇浓度而带来抗动脉粥样硬化的益处。巨噬细胞至粪便RCT方法在基因工程小鼠中的应用提供了证据,表明HDL的这一主要特性与动脉粥样硬化易感性的变化密切相关。本章详细介绍了目前用于测量巨噬细胞体外胆固醇流出或体内巨噬细胞特异性RCT的方法。本文所述的一般原则和技术可用于测量巨噬细胞培养物中人体血清的体外胆固醇流出能力,并评估不同实验病理生理条件的影响或不同治疗策略对小鼠体内巨噬细胞RCT调节的疗效。