INTRODUCTION: To test the hypothesis that administration of adenosine during reperfusion attenuates endothelial dysfunction and extension of infarct size by inhibiting polymorphonuclear neutrophil (PMN)-mediated events and apoptosis. METHODS: Anesthetized dogs were subjected to 1 h coronary artery occlusion and 6 h of reperfusion with infusion of saline (vehicle, n = 8) or 140 micrograms/kg per min adenosine, n = 8) continuously into the left atrium starting 5 min before reperfusion and continuing for 2 h. RESULTS: There was no intergroup difference in collateral myocardial blood flow measured by using colored microspheres in the area at risk during ischemia. Infusion of adenosine transiently improved segmental shortening (4.1 +/- 3.1% versus -2.5 +/- 2.3%, P < 0.05) and segmental work (41.4 +/- 22 versus 15 +/- 13 mmHg/mm, P < 0.05) after 4 h of reperfusion. Infusion of adenosine reduced size of infarct (determined by staining with triphenyltetrazolium chloride) from 27 +/- 2% with vehicle to 14 +/- 1%, (P < 0.05). This was confirmed by measuring that it lowered activity of plasma creatine kinase (from 19 +/- 2 versus 8 +/- 1 IU/g protein, P < 0.05). It also reduced the proportion of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling-positive nuclei in the perinecrotic zone from 17.3 +/- 1.6 to 10.3 +/- 1.0% (P < 0.05) and reduced the appearance of DNA ladders in gel electrophoresis. In addition, it significantly decreased accumulation of PMN in the ischemic area (determined by immunohistochemistry with anti-CD18 antibody) and activity of cardiac myeloperoxidase compared with vehicle (439 +/- 52 versus 183 +/- 20 PMN/mm2 myocardium and 1.1 +/- 0.1 versus 2.4 +/- 0.2 U/100 mg tissue, P < 0.05, respectively). Furthermore, infusion of adenosine during reperfusion preserved vascular endothelial function expressed in terms of a decrease in adherence of PMN to postischemic coronary artery endothelium (63 +/- 3 versus 36 +/- 4 PMN/mm2 endothelium, P < 0.05, basal function) and agonist (acetylcholine)-induced endothelium-dependent relaxation (negative logarithm to base 10 of concentration (mol/l) for half-maximal effect 7.7 +/- 0.1 versus 7.2 +/- 0.1, P < 0.05, stimulated function). Infusion of adenosine directly inhibited generation of superoxide radical from canine PMN in vitro dose dependently from 27.8 +/- 6.3 to 5.8 +/- 2.1 nmol/l/5 x 10(6) PMN (P < 0.05). CONCLUSION: Intra-atrial infusion of adenosine during reperfusion reduced accumulation of PMN in area at risk, preserved vascular endothelial function after ischemia-reperfusion by inhibiting interaction between PMN and endothelial cells, and decreased extension of infarct, possibly by limiting apoptosis.
INTRODUCTION: To test the hypothesis that administration of adenosine during reperfusion attenuates endothelial dysfunction and extension of infarct size by inhibiting polymorphonuclear neutrophil (PMN)-mediated events and apoptosis. METHODS: Anesthetized dogs were subjected to 1 h coronary artery occlusion and 6 h of reperfusion with infusion of saline (vehicle, n = 8) or 140 micrograms/kg per min adenosine, n = 8) continuously into the left atrium starting 5 min before reperfusion and continuing for 2 h. RESULTS: There was no intergroup difference in collateral myocardial blood flow measured by using colored microspheres in the area at risk during ischemia. Infusion of adenosine transiently improved segmental shortening (4.1 +/- 3.1% versus -2.5 +/- 2.3%, P < 0.05) and segmental work (41.4 +/- 22 versus 15 +/- 13 mmHg/mm, P < 0.05) after 4 h of reperfusion. Infusion of adenosine reduced size of infarct (determined by staining with triphenyltetrazolium chloride) from 27 +/- 2% with vehicle to 14 +/- 1%, (P < 0.05). This was confirmed by measuring that it lowered activity of plasma creatine kinase (from 19 +/- 2 versus 8 +/- 1 IU/g protein, P < 0.05). It also reduced the proportion of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling-positive nuclei in the perinecrotic zone from 17.3 +/- 1.6 to 10.3 +/- 1.0% (P < 0.05) and reduced the appearance of DNA ladders in gel electrophoresis. In addition, it significantly decreased accumulation of PMN in the ischemic area (determined by immunohistochemistry with anti-CD18 antibody) and activity of cardiac myeloperoxidase compared with vehicle (439 +/- 52 versus 183 +/- 20 PMN/mm2 myocardium and 1.1 +/- 0.1 versus 2.4 +/- 0.2 U/100 mg tissue, P < 0.05, respectively). Furthermore, infusion of adenosine during reperfusion preserved vascular endothelial function expressed in terms of a decrease in adherence of PMN to postischemic coronary artery endothelium (63 +/- 3 versus 36 +/- 4 PMN/mm2 endothelium, P < 0.05, basal function) and agonist (acetylcholine)-induced endothelium-dependent relaxation (negative logarithm to base 10 of concentration (mol/l) for half-maximal effect 7.7 +/- 0.1 versus 7.2 +/- 0.1, P < 0.05, stimulated function). Infusion of adenosine directly inhibited generation of superoxide radical from canine PMN in vitro dose dependently from 27.8 +/- 6.3 to 5.8 +/- 2.1 nmol/l/5 x 10(6) PMN (P < 0.05). CONCLUSION: Intra-atrial infusion of adenosine during reperfusion reduced accumulation of PMN in area at risk, preserved vascular endothelial function after ischemia-reperfusion by inhibiting interaction between PMN and endothelial cells, and decreased extension of infarct, possibly by limiting apoptosis.
Authors: Francisco Villarreal; Scott Zimmermann; Lala Makhsudova; Annika C Montag; Mark D Erion; David A Bullough; Bruce R Ito Journal: Mol Cell Biochem Date: 2003-09 Impact factor: 3.396