In vivo opening of the mitochondrial permeability transition pore in a rat model of ventricular fibrillation and closed-chest resuscitation.

Opening of the mitochondrial permeability transition pore (mPTP) is considered central to reperfusion injury. Yet, most of our knowledge comes from observations in isolated mitochondria, cells, and organs. We used a rat model of ventricular fibrillation (VF) and closed-chest resuscitation to examine whether the mPTP opens in vivo and whether cyclosporine A (CsA) attenuates the associated myocardial injury. Two series of 26 and 18 rats each underwent 10 minutes of untreated VF before attempting resuscitation. In series-1, rats received 50 µCi of tritium-labeled 2-deoxyglucose ([3H]DOG) harvesting their hearts at baseline (n=5), during VF (n=5), during resuscitation (n=6), and at post-resuscitation 60 minutes (n=5) and 240 minutes (n=5). mPTP opening was estimated measuring the ratio of mitochondria to left ventricular intracellular [3H]. In series-2, rats received 10 mg/kg of CsA or vehicle before resuscitation, measuring mitochondrial NAD+ content to indirectly assess mPTP opening. In Series-1, the mPTP opening ratio vs baseline (10.4 ± 1.9) increased during VF (16.8 ± 2.4, NS), closed-chest resuscitation (20.8 ± 6.3, P<0.05), and at post-resuscitation 60 minutes (20.9 ± 4.7, P<0.05) and 240 minutes (25.7 ± 11.0, P<0.01). In series 2, CsA failed to attenuate reductions in mitochondrial NAD+ and did not affect plasma cytochrome c, plasma cardiac troponin I, myocardial function, and survival. We report for the first time in an intact rat model of VF that mPTP opens during closed-chest resuscitation consistent with previous observations in mitochondria, cells, and organs of mPTP opening upon reperfusion. CsA, at the dose of 10 mg/kg neither prevented mPTP opening nor attenuated post-resuscitation myocardial injury.