Yamamoto Norio, Ueda Manabu, Sato Takuya, Kawasaki Kengo, Sawada Keisuke, Kawabata Kyuichi, Ashida Hitoshi
Food Science Research Center, House Wellness Foods Corporation, Hyogo, Japan.
Curr Protoc Pharmacol. 2011 Dec;Chapter 12:Unit 12.14.1-22. doi: 10.1002/0471141755.ph1214s55.
Facilitative glucose uptake transport systems are ubiquitous in animal cells and responsible for transporting glucose across the cell surface membrane. Evaluation of glucose uptake is crucial in the study of numerous diseases and metabolic disorders, such as myocardial ischemia, diabetes mellitus, and cancer. Methods for assessing glucose uptake into mammalian cells are detailed in this unit. The work is divided into four sections: (1) a brief overview of glucose uptake assays in cultured cells; (2) a method for measuring glucose uptake using radiolabeled 3-O-methylglucose; (3) a method for measuring glucose uptake using radiolabeled 2-deoxyglucose (2DG); and (4) an improved method for measuring 2DG-uptake using an enzymatic, fluorometric assay, eliminating the need for radiolabeled glucose analogs.
易化型葡萄糖摄取转运系统在动物细胞中普遍存在,负责将葡萄糖转运穿过细胞膜表面。葡萄糖摄取评估在众多疾病和代谢紊乱(如心肌缺血、糖尿病和癌症)的研究中至关重要。本单元详细介绍了评估哺乳动物细胞摄取葡萄糖的方法。这项工作分为四个部分:(1)培养细胞中葡萄糖摄取测定的简要概述;(2)使用放射性标记的3-O-甲基葡萄糖测量葡萄糖摄取的方法;(3)使用放射性标记的2-脱氧葡萄糖(2DG)测量葡萄糖摄取的方法;(4)一种使用酶促荧光测定法测量2DG摄取的改进方法,无需使用放射性标记的葡萄糖类似物。