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Protein kinase C- mediates adenosine A₃ receptor-induced delayed cardioprotection in mouse

Division of Cardiology, Department of Medicine, Medical College of Virginia, Virginia Commonwealth University, Richmond, Virginia 23298

Submitted 30 January 2003 ; accepted in final form 14 March 2003

We investigated the role of protein kinase C in adenosine A₃ receptor (A₃AR)-induced delayed cardioprotection in the mouse heart. Mice were treated with selective A₃AR agonist N⁶-(3-iodobenzyl)adenosine-5'-N-methyluronamide (IB-MECA). Twenty-four hours later, hearts were perfused in the Langendorff mode and subjected to 30 min of global ischemia and 30 min of reperfusion. Infarct size was determined by computer morphometry of tetrazolium-stained sections, and ventricular function was monitored by inserting a fluid-filled balloon into the left ventricle (LV). Chelerythrine chloride (CHE, 5.0 mg/kg) and rottlerin (Rot, 0.3 mg/kg) were given 30 min before IB-MECA to block total and PKC- isoforms, respectively. IB-MECA caused postischemic reduction in necrosis and improvement in ventricular function, which was abolished by CHE. Western blot analysis demonstrated translocation of the PKC- isoform but not the , , , isoform(s) from cytoplasm to the membrane fraction after 30 min of IB-MECA administration. A₃AR antagonist MRS-1191 and CHE blocked the translocation of PKC-. Furthermore, IB-MECA-induced increase in nuclear factor-B binding was diminished by CHE. These results provide direct evidence of an essential role of PKC, and more specifically, PKC- in A₃AR-induced delayed cardioprotection.

adenosine; nuclear factor -B; ischemia; reperfusion

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