Activation of protein kinase C increases adenosine production in the hypoxic canine coronary artery through the extracellular pathway.
Both ischemia and hypoxia increase adenosine production in the heart. This study tested whether hypoxia increases adenosine production in the coronary artery via ecto-5'-nucleotidase and the role of protein kinase C in this condition. Canine left circumflex coronary artery was rapidly removed and incubated in 10 mL Krebs-Henseleit solution for 30 minutes. The Krebs-Henseleit solution contained 5'-iodotubercidin and 2'-deoxycoformycin, which inhibit adenosine kinase and adenosine deaminase, respectively. Adenosine production was measured in intact coronary arteries under normoxic conditions (16.2 +/- 1.2 pmol/mg protein). Adenosine production was reduced by 27% after removal of endothelium. Ecto-5'-nucleotidase activity of coronary arteries with and without endothelium was 51 +/- 6 and 41 +/- 4 nmol/mg protein per minute under normoxic conditions. Hypoxia increased adenosine production to 27.0 +/- 2.3 and 20.0 +/- 0.8 pmol/mg protein with and without endothelium. Hypoxia also increased ecto-5'-nucleotidase activity of coronary arteries with and without endothelium (74 +/- 8 and 53 +/- 5 nmol/mg protein per minute; P < .05). Increases in adenosine production under hypoxic conditions were blunted by both an inhibitor of ecto-5'-nucleotidase and inhibitors of protein kinase C. Activation of ecto-5'-nucleotidase was blunted by an inhibitor of protein kinase C. These results indicate that hypoxia increased extracellular adenosine production and activated ecto-5'-nucleotidase via activation of protein kinase C in coronary arterial smooth muscle and endothelial cells. Increased adenosine production in coronary arteries during hypoxia may contribute to coronary vasodilation and cardioprotection against ischemic injury.
D015720:5'-Nucleotidase / Q000378:metabolism; D000241:Adenosine / Q000096:biosynthesis*; D058892:Adenosine Deaminase Inhibitors; D000248:Adenosine Kinase / Q000037:antagonists & inhibitors; D000818:Animals; D001158:Arteries / Q000378:metabolism; D003331:Coronary Vessels / Q000378:metabolism*; D004285:Dogs; D004789:Enzyme Activation; D000860:Hypoxia / Q000378:metabolism*; D066298:In Vitro Techniques; D015649:Pentostatin / Q000494:pharmacology; D011493:Protein Kinase C / Q000378:metabolism*; D014372:Tubercidin / Q000031:analogs & derivatives / Q000494:pharmacology
D058892:Adenosine Deaminase Inhibitors; C034214:5-iodotubercidin; D015649:Pentostatin; D000248:Adenosine Kinase; D011493:Protein Kinase C; D015720:5'-Nucleotidase; D000241:Adenosine; D014372:Tubercidin