Vesnarinone inhibits adenosine uptake in endothelial cells, smooth muscle cells and myocytes, and mediates cytoprotection

Vesnarinone is a novel synthetic inotropic agent. Recently, it has been reported that vesnarinone inhibits adenosine uptake in the B-lymphocytoid cell line. Since extracellular adenosine is cardioprotective, we examined whether vesnarinone inhibits adenosine uptake in cells constituting the cardiovascular system. 1 microCi of -3H-adenosine was added to cells of the myocyte cell line (C2C12), human coronary smooth muscle cell line (HCASMC), human and bovine coronary endothelial cell lines (HCAEC and BCAEC), bovine arterial endothelial cell line (BAEC), and human umbilical venous endothelial cell line (HUVEC). After 10 s-5 min, cells were separated from free [3H]adenosine, and the radioactivity was measured. When 0.1-100 microM of vesnarinone was added to each cell line, the uptake of adenosine was inhibited dose-dependently {% inhibition of -3H-adenosine uptake at 10 and 30 microM of vesnarinone: 14 and 33% (C2C12), 47 and 72% (HCASMC), 37 and 58% (HCAEC), 42 and 68% (BCAEC), 19 and 68% (BAEC), 29 and 59% (HUVEC)}. The cellular viability of HCAEC exposed to 60 min of hypoxia and 60 min of reoxygenation increased from 34+/-5 to 67+/-6% (Trypan blue exclusion test) and 23+/-5 to 78+/-6% (LDH release), which was completely blunted by 8-sulfophenyltheophylline, an adenosine receptor antagonist, and was partially blunted by alpha,beta-methyleneadenosine 5'-diphosphate, an inhibitor of ecto-5'-nucleotidase. We also found that vesnarinone is cytoprotective against hypoxia and reoxygenation in C2C12 and HCASMC. We conclude that vesnarinone inhibits the uptake of adenosine in cardiovascular cells, which contributes to cytoprotection.