Abstract
Mitochondrial permeability transition (MPT) is critical in cardiomyocyte death during reperfusion but it is not the only mechanism responsible for cell injury. The objectives of the study is to investigate the role of the duration of myocardial ischemia on mitochondrial integrity and cardiomyocyte death. Mitochondrial membrane potential (ΔΨm, JC-1) and MPT (calcein) were studied in cardiomyocytes from wild-type and cyclophilin D (CyD) KO mice refractory to MPT, submitted to simulated ischemia and 10 min reperfusion. Reperfusion after 15 min simulated ischemia induced a rapid recovery of ΔΨm, extreme cell shortening (contracture) and mitochondrial calcein release, and CyD ablation did not affect these changes or cell death. However, when reperfusion was performed after 25 min simulated ischemia, CyD ablation improved ΔΨm recovery and reduced calcein release and cell death (57.8 ± 4.9% vs. 77.3 ± 4.8%, P < 0.01). In a Langendorff system, CyD ablation increased infarct size after 30 min of ischemia (61.3 ± 6.4% vs. 45.3 ± 4.0%, P = 0.02) but reduced it when ischemia was prolonged to 60 min (52.8 ± 8.1% vs. 87.6 ± 3.7%, P < 0.01). NMR spectroscopy in rat hearts showed a rapid recovery of phosphocreatine after 30 min ischemia followed by a marked decay associated with contracture and LDH release, that were preventable with contractile blockade but not with cyclosporine A. In contrast, after 50 min ischemia, phosphocreatine recovery was impaired even with contractile blockade (65.2 ± 4% at 2 min), and cyclosporine A reduced contracture, LDH release and infarct size (52.1 ± 4.2% vs. 82.8 ± 3.6%, P < 0.01). In conclusion, the duration of ischemia critically determines the importance of MPT on reperfusion injury. Mechanisms other than MPT may play an important role in cell death after less severe ischemia.
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Acknowledgements
This study was supported by the Spanish Ministry of Science and Instituto de Salud Carlos III (RETICS-RECAVA RD06/0014/0025; CICYT SAF/2008-03067, FIS-PI080238 and PS09/02034). Ignasi Barba is recipient of a Ramon y Cajal fellowship.
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Supplementary Figure 1: Sequence of confocal images (60×) of 2 cardiac myocytes isolated from a wild-type mouse heart loaded with JC-1, under baseline conditions, after 15 min simulated ischemia, and during the first minutes of reperfusion. The graph corresponds to ΔΨm kinetics of these 2 cells throughout ischemia-reperfusion (arrow points the onset of reperfusion).
Supplementary Figure 2: Confocal images (60×) of a wild-type JC-1 loaded cardiac myocyte under baseline conditions, at 15 min of simulated ischemia, and after 3 min of reperfusion in the presence of the contractile blocker BDM (15 mmol/L). The graph corresponds to ΔΨm kinetics of this cell throughout 15 min ischemia-10 min reperfusion (arrow points the onset of reperfusion).
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Ruiz-Meana, M., Inserte, J., Fernandez-Sanz, C. et al. The role of mitochondrial permeability transition in reperfusion-induced cardiomyocyte death depends on the duration of ischemia. Basic Res Cardiol 106, 1259–1268 (2011). https://doi.org/10.1007/s00395-011-0225-5
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DOI: https://doi.org/10.1007/s00395-011-0225-5