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Involvement of NADPH oxidase in A2A adenosine receptor-mediated increase in coronary flow in isolated mouse hearts

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Abstract

Adenosine increases coronary flow mainly through the activation of A2A and A2B adenosine receptors (ARs). However, the mechanisms for the regulation of coronary flow are not fully understood. We previously demonstrated that adenosine-induced increase in coronary flow is in part through NADPH oxidase (Nox) activation, which is independent of activation of either A1 or A3ARs. In this study, we hypothesize that adenosine-mediated increase in coronary flow through Nox activation depends on A2A but not A2BARs. Functional studies were conducted using isolated Langendorff-perfused mouse hearts. Hydrogen peroxide (H2O2) production was measured in isolated coronary arteries from WT, A2AAR knockout (KO), and A2BAR KO mice using dichlorofluorescein immunofluorescence. Adenosine-induced concentration-dependent increase in coronary flow was attenuated by the specific Nox2 inhibitor gp91 ds-tat or reactive oxygen species (ROS) scavenger EUK134 in both WT and A2B but not A2AAR KO isolated hearts. Similarly, the A2AAR selective agonist CGS-21680-induced increase in coronary flow was significantly blunted by Nox2 inhibition in both WT and A2BAR KO, while the A2BAR selective agonist BAY 60-6583-induced increase in coronary flow was not affected by Nox2 inhibition in WT. In intact isolated coronary arteries, adenosine-induced (10 μM) increase in H2O2 formation in both WT and A2BAR KO mice was attenuated by Nox2 inhibition, whereas adenosine failed to increase H2O2 production in A2AAR KO mice. In conclusion, adenosine-induced increase in coronary flow is partially mediated by Nox2-derived H2O2, which critically depends upon the presence of A2AAR.

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Acknowledgments

The authors would like to thank Dr. Xueping Zhou for assistance with the immunofluorescence staining studies. This study was supported by NIH grants of HL027339, HL09444, HL071802, and U54GM104942.

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Authors and Affiliations

  1. Department of Physiology and Pharmacology, Center for Cardiovascular and Respiratory Sciences, Clinical and Translational Science Institute, West Virginia University, Morgantown, WV, 26506-9104, USA

    Zhichao Zhou, Uthra Rajamani, Hicham Labazi, Bunyen Teng & S. Jamal Mustafa

  2. Department of Medicine, University of North Carolina, Chapel Hill, NC, USA

    Stephen L. Tilley

  3. Universite Libre de Bruxelles, Brussels, Belgium

    Catherine Ledent

  4. Department of Pathology and Laboratory Medicine, University of California Davis Medical Centre, Sacramento, CA, USA

    Uthra Rajamani

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  1. Zhichao Zhou
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  2. Uthra Rajamani
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Correspondence to S. Jamal Mustafa.

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Zhou, Z., Rajamani, U., Labazi, H. et al. Involvement of NADPH oxidase in A2A adenosine receptor-mediated increase in coronary flow in isolated mouse hearts. Purinergic Signalling 11, 263–273 (2015). https://doi.org/10.1007/s11302-015-9451-x

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