Naked small interfering RNA of caspase-3 in preservation solution and autologous blood perfusate protects isolated ischemic porcine kidneys

Transplantation. 2011 Mar 15;91(5):501-7. doi: 10.1097/TP.0b013e318207949f.

Abstract

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 μg/ml) with the renal vein clamped and into autologous blood (0.15 μg/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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acid-Base Equilibrium
  • Animals
  • Apoptosis / drug effects
  • Caspase 3 / genetics*
  • Caspase 3 / metabolism
  • Caspase Inhibitors
  • Cryopreservation / methods*
  • Graft Survival
  • Kidney / drug effects
  • Kidney / metabolism
  • Kidney / pathology
  • Kidney Transplantation* / pathology
  • Models, Animal
  • Organ Preservation Solutions / pharmacology
  • Organ Preservation Solutions / therapeutic use*
  • Oxygen Consumption / drug effects
  • RNA, Small Interfering / pharmacology
  • RNA, Small Interfering / therapeutic use*
  • Reperfusion
  • Reperfusion Injury / metabolism
  • Reperfusion Injury / pathology
  • Reperfusion Injury / prevention & control*
  • Swine
  • Warm Ischemia

Substances

  • Caspase Inhibitors
  • Organ Preservation Solutions
  • RNA, Small Interfering
  • Caspase 3