engleski

Pulsatile Preservation Reduces Apoptosis in a Porcine Model of Kidney Donation after Cardiac Death (DCD)

The organ-donor shortage has led to greater use of DCD kidneys. Clinical studies suggest pulsatile preservation (PP) confers protection vs. traditional static preservation (SP). The mechanism of this protection is not known. The aim of this study was to determine whether PP reduced kidney cell death in a porcine model of DCD. We hypothesized that PP would result in 1) less apoptosis, 2) reduced activity of caspase-3, a major mediator of apoptosis, 3) activation of HIF-1α which is known to protect against apoptosis. Methods: 30 kg male pigs were subjected to myocardial ischemia and infarction. The supra-renal aorta was clamped and the infra-renal IVC vented. The pigs were exsanguinated and 1L of UW solution at 4°C was infused. The right kidney was removed and stored in cold KPS at 4°C for 24 hrs (SP). The contralateral kidney was perfused with cold KPS at 4°C for 24 hrs (PP) using a pulsatile perfusion device (Organ Recovery Systems, Illinois, USA). Apoptotic tubular cells/hpf were quantitated by a renal pathologist. Caspase-3/7 activity was measured using fluorescent substrate DEVD-AMC. HIF-1 activity was measured by ELISA (pg/10μg). Results: Apoptotic tubular cells/hpf were significantly greater at 24 hrs of SP vs. PP. Caspase-3/7 activity was not significantly different between SP and PP kidneys, but HIF activity was significantly increased in PP kidneys. Conclusion: We have shown that PP kidneys have significantly fewer apoptotic tubular cells/hpf than SP kidneys. This finding may explain the superior performance of PP vs. SP seen clinically. This protection is independent of caspase-3/7 activation, but is associated with HIF-1α activation at 24 hrs. of PP. Further study of the mechanisms by which pulsatile perfusion reduces apoptosis and increases HIF-1α activity is warranted.

transplantation; cold ischemia; apoptosis

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