Inhibition of adenosine re-uptake by nucleoside transport inhibitors, such as dipyridamole and dilazep, is proposed to increase extracellular levels of adenosine and thereby potentiate adenosine receptor-dependent pathways that promote cardiovascular health. Thus, adenosine can act as a paracrine and/or autocrine hormone which has been shown to regulate glucose uptake in some cell types. However, the role of adenosine in modulating glucose transport in cardiomyocytes is not clear. Therefore, we investigated whether exogenously applied adenosine, or inhibition of adenosine transport by nitrobenzylthioinosine (NBTI), dipyridamole or dilazep modulated basal and insulin-stimulated glucose uptake in the murine cardiomyocyte cell line, HL-1. HL-1 cell lysates were subjected to SDS-PAGE and immunoblotting to determine which GLUT isoforms are present. Glucose uptake was measured in the presence of dipyridamole (3-300 µM), dilazep (1-100 µM), NBTI (10-500 nM) and adenosine (50, 250 µM) or the non-metabolizable adenosine analog, 2-chloro-adenosine (250 µM). Our results demonstrated that HL-1 cells possess GLUT1 and GLUT4, the isoforms typically present in cardiomyocytes. We found no evidence for adenosine-dependent regulation of basal or insulin-stimulated glucose transport in HL-1 cardiomyocytes. However, we did observe a dose-dependent inhibition of glucose transport by dipyridamole (basal, IC₅₀= 12.2 µM, insulin-stimulated, IC₅₀= 13.09 µM) and dilazep (basal, IC₅₀= 5.7 µM, insulin-stimulated, IC₅₀= 19 µM) but not NBTI. Thus, our data suggest that dipyridamole and dilazep, which are widely used to specifically inhibit nucleoside transport, have a broader spectrum of transport inhibition than previously described. Moreover, these data may explain previous observations in which dipyridamole was noted to be pro-ischemic at high doses.