Metabolism of extracellular adenine nucleotides by human endothelial cells exposed to reactive oxygen metabolites

Abstract

Endothelial cells have ectonucleotidases that rapidly catabolize extracellular nucleotides. Our aim was to study whether the metabolism of extracellular nucleotides and adenosine are influenced by exposure of endothelial cells to reactive oxygen metabolites at concentrations relevant to human pathology. Human umbilical vein endothelial cells were incubated with hypoxanthine (100 microM) and xanthine oxidase (80 mU/ml), to generate superoxide, or with hydrogen peroxide (100 microM). The cells were then washed, and the metabolism of radioactive substrates was followed. After exposure to hypoxanthine-xanthine oxidase the half time of disappearance of [14C]ATP (5 microM) was prolonged from 9.9 +/- 5 to 28.3 +/- 15.6 min and that of [14C]AMP from 9.5 +/- 2.5 to 25.0 +/- 9.9 min. The conversion of extra- into intracellular nucleotides via adenosine was also decreased (mean for [14C]ATP 0.25 vs. 0.90 and for [14C]AMP, 0.075 vs. 0.75 nmol/10(6) cells in 30 min compared with parallel controls, respectively). Hydrogen peroxide or trypsin had no significant effect on the metabolism of extracellular adenine nucleotides and neither did a short (up to 15 min) exposure to the superoxide-generating system. The conversion of [14C]adenosine into intracellular nucleotides and hypoxanthine was not influenced by either hypoxanthine-xanthine oxidase or by hydrogen peroxide. We conclude that superoxide radicals inhibit the catabolism of extracellular adenine nucleotides by the ectonucleotidases of endothelial cells and may thus modify the pathophysiology of ischemia-reperfusion injury.