Roles of the unselective adenosine receptor agonist 5'-(N-ethylcarboxamido) adenosine-induced cardio-protection in vitro / 中国药学杂志

ABSTRACT

OBJECTIVE: To study the roles of adenosine A2A and A2B receptor in 5'-(N-ethylcarboxamido) adenosine (NECA)-induced cardioprotection in vitro against reperfusion injury, and to explore the underlying mechanism. METHODS: Simulated ischemia/reperfusion injury model was developed in cardiac H9c2 cells. NECA, an unselective adenosine receptor agonist, the selective antagonists of adenosine A2A receptor antagonist SCH58261 (SCH) and the selective A2B receptor antagonist MRS1706 (MRS) were used. CCK-8 assay was used to evaluate cell viability. Mitochondrial membrane potential (ΔΨm) was measured with fluorescence microscope using JC-1. Amplex Red Hydrogen Peroxide/Peroxidase Assay Kit was used to detect the level of intracellular H2O2. Intracellular reactive oxygen species (ROS) levels were determined with DCFH-DA. Mitochondrial ROS were detected with MitoSox Red. RESULTS: NECA applied at reperfusion reduced cell death in cells subjected to simulated ischemia/reperfusion. Compared with the ischemia/reperfusion injury group, NECA inhibited the reduction of cell viability and ΔΨm, and the elevation of intracellular and mitochondrial ROS, which were all abolished by adenosine A2A and A2B receptor antagonists(P < 0.01 or P < 0.05). CONCLUSION: Adenosine A2A and A2B receptors work in concert to mediate the cardioprotective effect of NECA presumably by modulating the mPTP opening and mitochondrial ROS generation.