Intermittent activation of bradykinin B2 receptors and mitochondrial KATP channels trigger cardiac postconditioning through redox signaling

Objective: Postconditioning (PostC) maneuvers allow post-ischemic accumulation of autacoids, which trigger protection. We tested if PostC-triggering includes bradykinin (BK) B-2 receptor activation and its downstream pathway. Methods and results: Isolated rat hearts underwent 30 min ischemia and 120 min reperfusion. Infarct size was evaluated using nitro-blue tetrazolium staining. In Control hearts infarct size was 61 +/- 5% of risk area. PostC (5 cycles of 10 s reperfusion/ischemia) reduced infarct size to 22 +/- 4% (p < 0.01). PostC protection was abolished by B-2 BK receptor-antagonists (HOE140 or WfN64338), nitric oxide synthase-inhibitor (L-nitro-arginine-methylester), protein kinase G (PKG)-blocker (8-bromoguanosine-3',5-cyclic-monophosphorothioate), and mitochondrial K-ATP (mK(ATP))-blocker (5-hydroxydecanoate) each given for 3 min only. Since 3 min of BK-infusion (100 nM) did not reproduce PostC protection, protocols with Intermittent-BK infusion were used to mimic PostC: a) 5 cycles of 10 s oxygenated-no-BK/ oxygenated + BK buffer; b) 5 cycles of 10 s oxygenated-no-BK/hypoxic + BK buffer. Both protocols with Intermittent-BK attenuated infarct size (36 +/- 5% and 38 +/- 4%, respectively; p < 0.05 vs Control and NS vs PostC for both; NS vs each other). Intermittent-BK protection was abolished by the same antagonists used to prevent PostC protection. Intermittence of re-oxygenation only (5 cycles of 10 s oxygenated/ hypoxic buffer) did not reproduce PostC. Yet, cardioprotection was triggered by intermittent mK(ATP) activation with diazoxide, but not by intermittent reactive oxygen species (ROS) generation with purine/xanthine oxidase. ROS scavengers (N-acetyl-L-cysteine or 2-mercaptopropionylglycine), given for 3 min only, abolished PostC-, Intermittent BK-and diazoxide-induced protection. Conclusions: Intermittent targeting of specific cellular sites (i.e. BK B-2 receptors and mK(ATP) channels) during early reperfusion triggers PostC protection via ROS signaling. Since neither intermittent oxygenation nor exogenous ROS generators can trigger protection, it is likely that intermittent autacoid accumulation and ROS compartmentalization may play a pivotal role in PostC-triggering. (c) 2007 European Society of Cardiology. Published by Elsevier B.V. All rights reserved.