Kener Kyle B, Munk Devin J, Hancock Chad R, Tessem Jeffery S
Nutrition, Dietetics and Food Science Department, Brigham Young University.
Nutrition, Dietetics and Food Science Department, Brigham Young University;
J Vis Exp. 2018 Jan 23(131):57053. doi: 10.3791/57053.
High-resolution respirometry allows for the measurement of oxygen consumption of isolated mitochondria, cells and tissues. Beta cells play a critical role in the body by controlling blood glucose levels through insulin secretion in response to elevated glucose concentrations. Insulin secretion is controlled by glucose metabolism and mitochondrial respiration. Therefore, measuring intact beta cell respiration is essential to be able to improve beta cell function as a treatment for diabetes. Using intact 832/13 INS-1 derived beta cells we can measure the effect of increasing glucose concentration on cellular respiration. This protocol allows us to measure beta cell respiration in the presence or absence of various compounds, allowing one to determine the effect of given compounds on intact cell respiration. Here we demonstrate the effect of two naturally occurring compounds, monomeric epicatechin and curcumin, on beta cell respiration under the presence of low (2.5 mM) or high glucose (16.7 mM) conditions. This technique can be used to determine the effect of various compounds on intact beta cell respiration in the presence of differing glucose concentrations.
高分辨率呼吸测定法可用于测量分离的线粒体、细胞和组织的耗氧量。β细胞在体内发挥着关键作用,它通过在血糖浓度升高时分泌胰岛素来控制血糖水平。胰岛素分泌受葡萄糖代谢和线粒体呼吸的控制。因此,测量完整β细胞的呼吸对于改善β细胞功能以治疗糖尿病至关重要。使用完整的源自832/13 INS-1的β细胞,我们可以测量葡萄糖浓度升高对细胞呼吸的影响。该方案使我们能够在存在或不存在各种化合物的情况下测量β细胞呼吸,从而确定给定化合物对完整细胞呼吸的影响。在此,我们展示了两种天然存在的化合物,单体表儿茶素和姜黄素,在低(2.5 mM)或高葡萄糖(16.7 mM)条件下对β细胞呼吸的影响。该技术可用于确定在不同葡萄糖浓度下各种化合物对完整β细胞呼吸的影响。