Journal
TRANSPLANTATION
Volume 91, Issue 5, Pages 501-507Publisher
LIPPINCOTT WILLIAMS & WILKINS
DOI: 10.1097/TP.0b013e318207949f
Keywords
Ischemia-reperfusion injury; Caspase-3; Apoptosis; Inflammation; Small interfering RNA; Isolated porcine kidneys
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Funding
- University Hospitals of Leicester NHS Trust
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Background. Ischemia-reperfusion injury (IRI) plays key roles in graft viability and posttransplantation function. Caspase-3 associated with apoptosis and inflammation is up-regulated by IRI. We propose that naked small interfering RNA (siRNA) targeting caspase-3 may prevent IRI and improve kidney preservation. Methods. Porcine kidneys were retrieved after 10-min warm ischemia and flushed with 500 mL hyperosmolar citrate. Caspase-3 siRNA was administered directly into the renal artery in hyperosmolar citrate solution (3 mu g/ml) with the renal vein clamped and into autologous blood (0.15 mu g/ml), both of which were stored for 24 hr on ice. The kidneys were reperfused with oxygenated autologous blood for 3 hr on an isolated organ perfusion system to assess renal function. Results. In the siRNA-treated group, the expression of 32-kDa caspase-3 precursor and 17-kDa active subunit was down-regulated by 30% and 56%, respectively, with 40% reduction in active caspase-3+ cells. Consequently, apoptotic cells in the renal cortex were significantly decreased by 47%, whereas inflammatory cells were only marginally reduced by 26%. Caspase-3 siRNA also doubled oxygen consumption and significantly neutralized perfusate pH. Renal blood flow was gradually increased from 5 min to 3 hr by caspase-3 siRNA and marginally improved at the end of 3-hr reperfusion by 45%. Conclusions. The administration of caspase-3 siRNA directly into the kidney and hemoperfusate during cold preservation improved IRI with reduced caspase-3 and apoptosis, and better renal oxygenation and acid-base homeostasis. This proof of principle experiment using a novel siRNA therapy in large animal organs provides valuable data for human kidney transplantation.
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