Zou Chenhui, Wang Yajie, Shen Zhufang
Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, 1 Xiannongtan Street, Beijing 100050, China.
J Biochem Biophys Methods. 2005 Sep 30;64(3):207-15. doi: 10.1016/j.jbbm.2005.08.001.
Evaluation of glucose uptake ability in cells plays a fundamental role in diabetes mellitus research. In this study, we describe a sensitive and non-radioactive assay for direct and rapid measuring glucose uptake in single, living cells. The assay is based on direct incubation of mammalian cells with a fluorescent d-glucose analog 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-2-deoxy-D-glucose (2-NBDG) followed by flow cytometric detection of fluorescence produced by the cells. A series of experiments were conducted to define optimal conditions for this assay. By this technique, it was found that insulin lost its physiological effects on cells in vitro meanwhile some other anti-diabetic drugs facilitated the cell glucose uptake rates with mechanisms which likely to be different from those of insulin or those that were generally accepted of each drug. Our findings show that this technology has potential for applications in both medicine and research.
细胞葡萄糖摄取能力的评估在糖尿病研究中起着基础性作用。在本研究中，我们描述了一种灵敏且无放射性的检测方法，用于直接快速测量单个活细胞中的葡萄糖摄取。该检测方法基于将哺乳动物细胞与荧光D - 葡萄糖类似物2 - [N - (7 - 硝基苯并 - 2 - 恶唑 - 1,3 - 二氮杂环丁烷 - 4 - 基)氨基] - 2 - 脱氧 - D - 葡萄糖（2 - NBDG）直接孵育，随后通过流式细胞术检测细胞产生的荧光。进行了一系列实验以确定该检测方法的最佳条件。通过该技术发现，胰岛素在体外对细胞失去了其生理作用，同时一些其他抗糖尿病药物促进了细胞葡萄糖摄取率，其机制可能与胰岛素不同，或者与每种药物通常被认可的机制不同。我们的研究结果表明，该技术在医学和研究领域均具有应用潜力。
