Measurement of Intestinal and Peripheral Cholesterol Fluxes by a Dual‐Tracer Balance Method

Q1 Agricultural and Biological Sciences

Current protocols in mouse biology Pub Date : 2016-12-01 DOI:10.1002/cpmo.16

O. Ronda, Theo H. Dijk, H. Verkade, A. Groen

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引用次数: 18

Abstract

Long‐term elevated plasma cholesterol levels put individuals at risk for developing atherosclerosis. Plasma cholesterol levels are determined by the balance between cholesterol input and output fluxes. Here we describe in detail the methodology to determine the different cholesterol fluxes in mice. The percentage of absorbed cholesterol is calculated from a stable isotope–based double‐label method. Cholesterol synthesis is calculated from MIDA after 13C‐acetate enrichment. Cholesterol is removed from the body via the feces. The fecal excretion route is either biliary or non‐biliary. The non‐biliary route is dominated by trans‐intestinal cholesterol efflux, or TICE. Biliary excretion of cholesterol is measured by collecting bile. Non‐biliary excretion is calculated by computational modeling. In this article, we describe methods and procedures to measure and calculate dietary intake of cholesterol, fractional cholesterol absorption, fecal neutral sterol output, biliary cholesterol excretion, TICE, cholesterol synthesis, peripheral fluxes, and whole‐body cholesterol balance. © 2016 by John Wiley & Sons, Inc.

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双示踪平衡法测量肠道和外周胆固醇通量

长期升高的血浆胆固醇水平使个体有发生动脉粥样硬化的风险。血浆胆固醇水平是由胆固醇输入和输出通量之间的平衡决定的。在这里，我们详细描述了确定小鼠不同胆固醇通量的方法。吸收胆固醇的百分比是通过基于稳定同位素的双标签方法计算的。13C‐acetate富集后，通过MIDA计算胆固醇合成。胆固醇通过粪便从体内排出。粪便的排泄途径为胆道或非胆道。非胆道途径主要是经肠胆固醇外排(TICE)。胆中胆固醇的排泄是通过收集胆汁来测定的。非胆汁排泄通过计算模型计算。在本文中，我们描述了测量和计算膳食胆固醇摄入量、部分胆固醇吸收、粪便中性胆固醇输出、胆汁胆固醇排泄、TICE、胆固醇合成、外周通量和全身胆固醇平衡的方法和程序。©2016 by John Wiley & Sons, Inc。

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Current protocols in mouse biology Agricultural and Biological Sciences-Animal Science and Zoology

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期刊介绍： Sound and reproducible laboratory methods are the foundation of scientific discovery. Yet, all too often, nuances that are critical for an experiment''s success are not captured in the primary literature but exist only as part of a lab''s oral tradition. The aim of Current Protocols in Mouse Biology is to provide the clearest, most detailed and reliable step-by-step instructions for protocols involving the use of mice in biomedical research. Written by experts in the field and extensively edited to our exacting standards, the protocols include all of the information necessary to complete an experiment in the laboratory—introduction, materials lists with supplier information, detailed step-by-step procedures with helpful annotations, recipes for reagents and solutions, illustrative figures and information-packed tables. Each article also provides invaluable discussions of background information, applications of the methods, important assumptions, key parameters, time considerations, and tips to help avoid common pitfalls and troubleshoot experiments. Furthermore, Current Protocols in Mouse Biology content is thoughtfully organized by topic for optimal usage and to maximize contextual knowledge. Quarterly issues allow Current Protocols to constantly evolve to keep pace with the newest discoveries and developments. Current Protocols in Mouse Biology brings together resources in mouse biology and genetics and provides a mouse protocol resource that covers all aspects of mouse biology. Current Protocols in Mouse Biology also permits optimization of mouse model usage, which is significantly impacted by both cost and ethical constraints. Optimal and standardized mouse protocols ultimately reduce experimental variability and reduce the number of animals used in mouse experiments.