Loss of ectonucleotidases from the coronary vascular bed after ischemia-reperfusion in isolated rat heart

BMC Cardiovascular Disorders

Isao Matsuoka³  Masa-aki Ito³  Junko Kimura¹  Masahiro Murakawa²  Kaoru Takahashi-Sato² 

[1] Departments of Pharmacology, Fukushima Medical University School of Medicine, Fukushima 960-1295, Japan;Departments of Anesthesiology, Fukushima Medical University School of Medicine, Fukushima 960-1295, Japan;Laboratory of Pharmacology, Faculty of Pharmacy, Takasaki University of Health and Welfare, Gunma 370-0033, Japan

关键词: Adenosine;    ATP;    Ectonucleotidase;    Coronary circulation;    Ischemia-reperfusion;   

Others  :  857577
DOI  :  10.1186/1471-2261-13-53

 received in 2012-11-26, accepted in 2013-07-25,  发布年份 2013

【 摘 要 】

Background

Ectonucleotidase plays an important role in the regulation of cardiac function by controlling extracellular levels of adenine nucleotides and adenosine. To determine the influence of ischemia-reperfusion injury on ectonucleotidase activity in coronary vascular bed, we compared the metabolic profile of adenine nucleotides during the coronary circulation in pre- and post-ischemic heart.

Methods

Langendorff-perfused rat hearts were used to assess the intracoronary metabolism of adenine nucleotides. The effects of ischemia on the adenine nucleotide metabolism were examined after 30 min of ischemia and 30 min of reperfusion. Adenine nucleotide metabolites were measured by high performance liquid chromatography.

Results

ATP, ADP and AMP were rapidly metabolized to adenosine and inosine during the coronary circulation. After ischemia, ectonucleotidase activity of the coronary vascular bed was significantly decreased. In addition, the perfusate from the ischemic heart contained a considerable amount of enzymes degrading ATP, AMP and adenosine. Immunoblot analysis revealed that the perfusate from the ischemic heart dominantly contained ectonucleoside triphosphate diphosphohydrolase 1, and, to a lesser extent, ecto-5’-nucleotidase. The leakage of nucleotide metabolizing enzymes from the coronary vascular bed by ischemia-reperfusion was more remarkable in aged rats, in which post-ischemic cardiac dysfunction was more serious.

Conclusion

Ectonucleotidases were liberated from the coronary vascular bed by ischemia-reperfusion, resulting in an overall decrease in ectonucleotidase activity in the post-ischemic coronary vascular bed. These results suggest that decreased ectonucleotidase activity by ischemia may exacerbate subsequent reperfusion injury, and that levels of circulating ectonucleotidase may reflect the severity of ischemic vascular injury.

【 授权许可】

   
2013 Takahashi-Sato et al.; licensee BioMed Central Ltd.

【 图 表 】

【 参考文献 】

• [1]Mubagwa K, Flameng W: Adenosine, adenosine receptors and myocardial protection: An updated overview. Cardiovasc Res 2001, 52:25-39.
• [2]Vassort G: Adenosine 5’-Triphosphate: a P2-purinergic agonist in the myocardium. Physiol Rev 2001, 81:767-806.
• [3]Korchazhkina O, Wright G, Exley C: Intravascular ATP and coronary vasodilation in the isolated working rat heart. Br J Pharmacol 1999, 127:701-708.
• [4]Burnstock G: Integration of factors controlling vascular tone: overview. Anesthesiology 1993, 79:1368-1380.
• [5]Lazarowski ER, Boucher RC, Harden TK: Mechanisms of release of nucleotides and integration of their action as P2X- and P2Y-receptor activating molecules. Mol Pharmacol 2003, 64:785-795.
• [6]Zimmermann H: Extracellular metabolism of ATP and other nucleotides. Naunyn Schmiedebergs Arch Pharmacol 2000, 362:299-309.
• [7]Enjyoji K, Sevigny J, Lin Y, Frenette PS, Christie PD, Esch JSA, Imai M, Edelberg JM, Rayburn H, Lech M, et al.: Targeted disruption of cd39/ATP diphosphohydrolase results in disordered hemostasis and thromboregulation. Nat Med 1999, 5:1010-1017.
• [8]Kaczmarek E, Koziak K, Sévigny J, Siegel JB, Anrather J, Beaudoin AR, Bach FH, Robson SC: Identification and characterization of CD39/vascular ATP diphosphohydrolase. J Biol Chem 1996, 271:33116-33122.
• [9]Mistry G, Drummond GI: Adenosine metabolism in microvessels from heart and brain. J Mol Cell Cardiol 1986, 18:13-22.
• [10]Eltzschig HK, Köhler D, Eckle T, Kong T, Robson SC, Colgan SP: Central role of Sp1-regulated CD39 in hypoxia/ischemia protection. Blood 2009, 113:224-232.
• [11]Kitakaze M, Hori M, Takashima S, Sato H, Inoue M, Kamada T: Ischemic preconditioning increases adenosine release and 5’-nucleotidase activity during myocardial ischemia and reperfusion in dogs. Implications for myocardial salvage. Circulation 1993, 87:208-215.
• [12]Robson SC, Kaczmarek E, Siegel JB, Candinas D, Koziak K, Millan M, Hancock WW, Bach FH: Loss of ATP diphosphohydrolase activity with endothelial cell activation. J Exp Med 1997, 185:153-164.
• [13]Yegutkin G, Helenius M, Kaczmarek E, Burns N, Jalkanen S, Stenmark K, Gerasimovskaya E: Chronic hypoxia impairs extracellular nucleotide metabolism and barrier function in pulmonary artery vasa vasorum endothelial cells. Angiogenesis 2011, 14:503-513.
• [14]Todorov LD, Mihaylova-Todorova S, Westfall TD, Sneddon P, Kennedy C, Bjur RA, Westfall DP: Neuronal release of soluble nucleotidases and their role in neurotransmitter inactivation. Nature 1997, 387:76-79.
• [15]Matsuoka I, Zhou Q, Ishimoto H, Nakanishi H: Extracellular ATP stimulates adenylyl cyclase and phospholipase C through distinct purinoceptors in NG108-15 cells. Mol Pharmacol 1995, 47:855-862.
• [16]Ohkubo S, Kimura J, Matsuoka I: Ecto-alkaline phosphatase in NG108-15 cells: a key enzyme mediating P1 antagonist-sensitive ATP response. Br J Pharmacol 2000, 131:1667-1672.
• [17]Candinas D, Koyamada N, Miyatake T, Siege IJ, Hancock WW, Bach FH, Robson SC: Loss of rat glomerular ATP diphosphohydrolase activity during reperfusion injury is associated with oxidative stress reactions. Thromb Haemost 1996, 76:807-812.
• [18]Braun N, Lenz C, Gillardon F, Zimmermann M, Zimmermann H: Focal cerebral ischemia enhances glial expression of ecto-5’ -nucleotidase. Brain Res 1997, 766:213-226.
• [19]Braun N, Zhu Y, Krieglstein J, Culmsee C, Zimmermann H: Upregulation of the Enzyme Chain Hydrolyzing Extracellular ATP after Transient Forebrain Ischemia in the Rat. J Neurosci 1998, 18:4891-4900.
• [20]Siegfried G, Amiel C, Friedlander G: Inhibition of Ecto-5’-nucleotidase by Nitric Oxide Donors: Implications in renal epithelial cells. J Biol Chem 1996, 271:4659-4664.
• [21]Kalsi K, Lawson C, Dominguez M, Taylor P, Yacoub M, Smolenski R: Regulation of ecto-5’-nucleotidase by TNF-alpha in human endothelial cells. Mol Cell Biochem 2002, 232:113-119.
• [22]Zinchuk V, Okada T, Seguchi H: Lipopolysaccharide alters ecto-ATP-diphosphohydrolase and causes relocation of its reaction product in experimental intrahepatic cholestasis. Cell Tissue Res 2001, 304:103-109.
• [23]Darvish A, Pomerantz RW, Zografides PG, Metting PJ: Contribution of cytosolic and membrane-bound 5’-nucleotidases to cardiac adenosine production. Am J Physiol Heart Circ Physiol 1996, 271:H2162-H2167.
• [24]Harris NR, Langlois KW: Age-dependent responses of the mesenteric vascular bed to ischemia-reperfusion. Am J Physiol Heart Circ Physiol 1998, 274:H1509-H1515.
• [25]Schulman D, Latchman DS, Yellon DM: Effect of aging on the ability of preconditioning to protect rat hearts from ischemia-reperfusion injury. Am J Physiol Heart Circ Physiol 2001, 281:H1630-H1636.
• [26]Bo¨nner F, Borg N, Burghoff S, Schrader J: Resident Cardiac Immune Cells and Expression of the Ectonucleotidase Enzymes CD39 and CD73 after Ischemic Injury. PLoS ONE 2012, 7:e34730.