Ko Chih-Wei, Qu Jie, Liu Min, Black Dennis D, Tso Patrick
Department of Pathology and Laboratory Medicine, University of Cincinnati, Cincinnati, Ohio.
Children's Foundation Research Institute at Le Bonheur Children's Hospital, University of Tennessee Health Science Center, Memphis, Tennessee.
Curr Protoc Mouse Biol. 2019 Mar;9(1):e60. doi: 10.1002/cpmo.60. Epub 2019 Feb 23.
This protocol provides a comprehensive reference for the evolution of the lymph fistula model, the mechanism of lipid absorption, the detailed procedure for studying lipid absorption using the lymph fistula model, the interpretation of the results, and consideration of the experimental design. The lymph fistula model is an approach to assess the concentration and rate of a range of molecules transported by the lymph by cannulating lymph duct in animals. In this protocol, mice first undergo surgery with the implantation of cannulae in the duodenum and mesenteric lymph duct and are allowed to recover overnight in Bollman restraining cages housed in a temperature-regulated environment. To study in vivo lipid absorption, a lipid emulsion is prepared with labeled tracers, including [ H]-triolein and [ C]-cholesterol. On the day of the experiment, mice are continuously infused with lipid emulsion via the duodenum for 6 hr, and lymph is usually collected hourly. At the end of the study, gastrointestinal segments and their luminal contents are collected separately for determination of the digestion, uptake, and transport of exogenous lipids. © 2019 by John Wiley & Sons, Inc.
本方案为淋巴瘘模型的演变、脂质吸收机制、使用淋巴瘘模型研究脂质吸收的详细步骤、结果解读以及实验设计考量提供了全面参考。淋巴瘘模型是一种通过在动物身上插管淋巴管来评估一系列分子经淋巴转运的浓度和速率的方法。在本方案中，小鼠首先接受手术，在十二指肠和肠系膜淋巴管中植入插管，然后在温度可控环境中的博尔曼约束笼中过夜恢复。为研究体内脂质吸收，制备含标记示踪剂的脂质乳剂，包括[H]-三油酸甘油酯和[C]-胆固醇。在实验当天，小鼠经十二指肠持续输注脂质乳剂6小时，通常每小时收集一次淋巴。在研究结束时，分别收集胃肠道段及其管腔内容物，以测定外源性脂质的消化、摄取和转运。© 2019约翰威立国际出版公司
