A(2B) adenosine receptors stimulate growth of porcine and rat arterial endothelial cells

The goal of this study was to determine which adenosine receptor subtype mediates growth stimulation by adenosine in arterial endothelial cells. In porcine coronary artery and rat aortic endothelial cells, 2-chloroadenosine (Cl-Ad), a metabolically stable analog of adenosine, stimulated DNA synthesis ((3)H-thymidine incorporation), cellular proliferation (cell number), collagen synthesis ((3)H-proline incorporation), and cell migration. The growth effects of adenosine and Cl-Ad were mimicked by the adenosine receptor agonist 5'-N-methylcarboxamidoadenosine but not by the adenosine receptor agonists N(6)-cyclopentyladenosine, 4-aminobenzyl-5'-N-methylcarboxamidoadenosine or CGS21680, an agonist profile consistent with an A(2B) receptor-mediated effect. The adenosine receptor antagonists KF17837 and 1,3-dipropyl-8-p-sulfophenylxanthine but not 8-cyclopentyl-1,3-dipropylxanthine blocked the growth-stimulatory effects of Cl-Ad and 5'-N-methylcarboxamidoadenosine, an antagonist profile consistent with an A(2) receptor-mediated action. Treatment of endothelial cells with erythro-9-(2-hydroxy-3-nonyl) adenine plus iodotubericidin (inhibitors of adenosine deaminase and adenosine kinase, respectively) induced endothelial cell growth, and these effects were blocked by 1,3-dipropyl-8-p-sulfophenylxanthine and KF17837 but not 8-cyclopentyl-1,3-dipropylxanthine, suggesting that endothelial cell-derived adenosine induces growth via A(2) receptors. The growth-stimulatory effects of Cl-Ad, 5'-N-methylcarboxamidoadenosine, and erythro-9-(2-hydroxy-3-nonyl) adenine plus iodotubericidin were abolished by antisense but not scrambled or sense oligonucleotides to the A(2B) receptor. Our findings strongly support the hypothesis that adenosine induces endothelial cell growth by activating A(2B) receptors. Thus, A(2B) receptors may play a critical role in regulating vascular remodeling associated with endothelial cell proliferation in angiogenesis, collateral vessel development, and recovery after vascular injury. Pharmacological or molecular biological activation of A(2B) receptors may be useful in modulating vascular remodeling.