ABSTRACT
Injury due to cold ischemia reperfusion (I/R) is a major cause of primary graft non-function following liver transplantation. We postulated that I/R-induced cellular damage during liver transplantation might affect the secretory pathway, particularly at the endoplasmic reticulum (ER). We examined the involvement of ER stress in organ preservation, and compared cold storage in University of Wisconsin (UW) solution and in Institute Georges Lopez-1 (IGL-1) solution. In one group of rats, livers were preserved in UW solution for 8 h at 4 °C, and then orthotopic liver transplantation was performed according to Kamada's cuff technique. In another group, livers were preserved in IGL-1 solution. The effect of each preservation solution on the induction of ER stress, hepatic injury, mitochondrial damage and cell death was evaluated. As expected, we found increased ER stress after liver transplantation. IGL-1 solution significantly attenuated ER damage by reducing the activation of three pathways of unfolded protein response and their effector molecules caspase-12, C/EBP homologous protein-10, X-box-binding protein 1, tumor necrosis factor-associated factor 2 and eukaryotic translation initiation factor 2. This attenuation of ER stress was associated with a reduction in hepatic injury and cell death. Our results show that IGL-1 solution may be a useful means to circumvent excessive ER stress reactions associated with liver transplantation, and may optimize graft quality.
Subject(s)
Liver Transplantation , Liver/metabolism , Organ Preservation Solutions/pharmacology , Organ Preservation/methods , Signal Transduction/drug effects , Adenosine/pharmacology , Allopurinol/pharmacology , Animals , Apoptosis/drug effects , Caspase 12/genetics , Caspase 12/metabolism , Cold Ischemia , Cold Temperature , DNA-Binding Proteins/genetics , DNA-Binding Proteins/metabolism , Endoplasmic Reticulum Stress/drug effects , Gene Expression/drug effects , Glutathione/pharmacology , Insulin/pharmacology , Liver/pathology , Male , Raffinose/pharmacology , Rats , Regulatory Factor X Transcription Factors , Reperfusion Injury/metabolism , Reperfusion Injury/pathology , Reperfusion Injury/prevention & control , Signal Transduction/genetics , TNF Receptor-Associated Factor 2/genetics , TNF Receptor-Associated Factor 2/metabolism , Transcription Factors/genetics , Transcription Factors/metabolism , Unfolded Protein Response/drug effects , Unfolded Protein Response/geneticsABSTRACT
University of Wisconsin (UW) preservation solution is considered an effective flush and cold storage liquid. However, recent studies have provided evidence of the hyperaggregating effect on human red blood cells (RBC) of hydroxyethyl starch (HES), one of the components of the UW solution. In contrast, preservation solutions containing polyethylene glycol (PEG) have been found to be effective for organ preservation. The aim of this study was to compare the effects of HES (50 g/L); PEG 20 kDa (50 and 30 g/L), and PEG35 kDa (1.05 g/L) added to UW on the rheologic parameters of human RBC at 4 degrees C. Sedimentation rate was measured by the Westergren procedure and blood viscosity evaluated at high shear rates using a cone/plate viscometer. Alterations in RBC morphology and aggregation were evaluated by light microscopy. RBC sedimentation and viscosity were not affected by the inversion of Na+ and K+ concentrations in UW, but were increased by HES. PEGs appeared to reduce RBC deformability with concomitant inhibition of RBC aggregation. These results were consistent with reduced viscosity for PEG-containing solutions. In conclusion, the use of PEG did not change the physiologic function of human RBCs and thus may be an alternative to HES in UW liquids.
Subject(s)
Blood Viscosity/drug effects , Erythrocyte Aggregation/drug effects , Hydroxyethyl Starch Derivatives/pharmacology , Organ Preservation Solutions/pharmacology , Polyethylene Glycols/pharmacology , Adenosine/pharmacology , Allopurinol/pharmacology , Erythrocyte Deformability/drug effects , Glutathione/pharmacology , Humans , Insulin/pharmacology , Potassium/pharmacology , Raffinose/pharmacology , Shear Strength , Sodium/pharmacology , Stress, MechanicalABSTRACT
We used the isolated perfused rat liver model (IPRL) to assess parenchymal and nonparenchymal cell integrity after different conditions of storage and reperfusion. Two studies were performed. In study 1, the IPRL was applied to evaluate the effects of 30 min of normothermic reperfusion with Elohes solution, enriched William's medium (Wif), or Carolina rinse solution (CRS) following 24 h of cold preservation in high-K+ or high-Na+ UW solution. As indicated by creatine kinase-BB (CK-BB) release, reperfusion with CRS provided greater protection of endothelial cells after storage in high-K+ UW solution than after storage in high-Na+ UW solution. In study 2, livers were cold-preserved (24 h, 4 degrees C) in either high-K+ or high-Na+ UW solution, then flushed with either CRS or Wif solution at room temperature before reperfusion (120 min, 37 degrees C) with 5% albumin-William's medium E. There was no statistical difference between the rinse solutions for bile flow and transaminases release. However, CRS improved bile indocyanine green excretion, which is known to be a marker of parenchymal and nonparenchymal cell integrity. Therefore, we can assume that this rinse solution protects rat liver grafts from reperfusion-induced microvascular damage.
