A fluorescent compound for glucose uptake measurements in isolated rat cardiomyocytes

Abstract

A focus of current diabetes research is the development of insulinomimetic compounds for oral treatment of diabetes and its associated cardiac complications. Screening compounds for their potential insulinomimetic effects usually involves the use of radioactive isotopes. The focus of this study was to investigate a nonradioactive fluorescent compound for its use in screening insulinomimetic compounds. The indicator 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose (2-NBDG) has been used by some workers to measure glucose uptake in Escherichia coli and Candida albicans. We propose that 2-NBDG will also be a suitable indicator for mammalian cell lines, in particular rat cardiomyocytes. We found that the indicator could give a reliable reproducible standard curve following appropriate dilution and is taken up by isolated cardiomyocytes. The insulinomimetic compounds vanadyl sulfate and sodium molybdate showed rates of glucose uptake similar to that of insulin. Furthermore, the rate of uptake measured for insulin using this technique (0.04 ± 0.003 nmol·min–1·106 cells–1) is comparable with previous literature using 2-deoxyglucose uptake measurements on isolated myocytes (0.040 nmol·min–1·106 cells–1), demonstrating the validity of this fluorescent compound for glucose uptake studies.Key words: cardiomyocytes, fluorescent deoxyglucose, glucose uptake, insulin mimics.