Activation of protein kinase B by adenosine A1 and A3 receptors in newborn rat cardiomyocytes

Renée Germack, Martin Griffin, John M. Dickenson

Research output: Contribution to journalArticle

Abstract

It is well established that adenosine receptors are involved in cardioprotection and that protein kinase B (PKB) is associated with cell survival. Therefore, in this study we have investigated whether adenosine receptors (A1, A2A and A3) activate PKB by Western blotting and determined the involvement of phosphatidylinositol 3-kinase (PI-3K)/PKB in adenosine-induced preconditioning in cultured newborn rat cardiomyocytes. Adenosine (non-selective agonist), CPA (A1 selective agonist) and Cl-IB-MECA (A(3) selective agonist) all increased PKB phosphorylation in a time- and concentration-dependent manner. The combined maximal response to CPA and Cl-IB-MECA was similar to the increase in PKB phosphorylation induced by adenosine alone. CGS 21680 (A2A selective agonist) did not stimulate an increase in PKB phosphorylation. Adenosine, CPA and Cl-IB-MECA-mediated PKB phosphorylation were inhibited by pertussis toxin (PTX blocks G(i)/G(o)-protein), genistein (tyrosine kinase inhibitor), PP2 (Src tyrosine kinase inhibitor) and by the epidermal growth factor (EGF) receptor tyrosine kinase inhibitor AG 1478. The PI-3K inhibitors wortmannin and LY 294002 blocked A(1) and A(3) receptor-mediated PKB phosphorylation. The role of PI-3K/PKB in adenosine-induced preconditioning was assessed by monitoring Caspase 3 activity and lactate dehydrogenase (LDH) release induced by exposure of cardiomyocytes to 4 h hypoxia (0.5% O2) followed by 18 h reoxygenation (HX4/R). Pre-treatment with wortmannin had no significant effect on the ability of adenosine-induced preconditioning to reduce the release of LDH or Caspase 3 activation following HX4/R. In conclusion, we have shown for the first time that adenosine A1 and A3 receptors trigger increases in PKB phosphorylation in rat cardiomyocytes via a G1/G0-protein and tyrosine kinase-dependent pathway. However, the PI-3K/PKB pathway does not appear to be involved in adenosine-induced cardioprotection by preconditioning

Adenosine A1 receptor
.

Original languageEnglish
Pages (from-to)989-999
Number of pages11
JournalJournal of Molecular and Cellular Cardiology
Volume37
Issue number5
DOIs
Publication statusPublished - Nov 2004

Keywords

  • Adenosine A1 receptor
  • adenosine A3 receptor
  • cardiomyocytes
  • protein kinase B
  • akt
  • hypoxia
  • ischaemia
  • reoxygenation
  • caspase 3
  • lactate dehydrogenase

Fingerprint Dive into the research topics of 'Activation of protein kinase B by adenosine A<sub>1</sub> and A<sub>3</sub> receptors in newborn rat cardiomyocytes'. Together they form a unique fingerprint.

  • Cite this