Normothermic Machine Perfusion Protects Against Liver Ischemia-Reperfusion Injury During Reduced-Size Liver Transplantation in Pigs

Abstract

BACKGROUND Normothermic machine perfusion (NMP) preservation is superior to cold preservation during reduced-size liver transplantation (RSLT) in pigs. However, the mechanism of this protective effect has not been explained. We aimed to compare the effects of NMP preservation with that of cold preservation (CS) in protecting against ischemia-reperfusion injury (IRI) during RSLT in pigs. MATERIAL AND METHODS Twenty-four healthy Bama miniature pigs were randomized into 2 groups: 1) the NMP group in which donor livers harvested without warm ischemia time and cardiac activity were connected to the NMP system to reduce liver size under normothermic conditions, and 2) the CS group in which donor livers harvested without warm ischemia time and cardiac activity were perfused using the University of Wisconsin (UW) solution and then preserved in the 0-4°C UW solution to reduce liver size under cold conditions. Livers were then transplanted without veno-venous bypass. Amounts of bile secretion for the NMP groups were recorded hourly. The serological indices were measured. Expressions of cytochrome C, caspase 3, and NF-κB p65 in liver tissue were observed. RESULTS The levels of bile secretions were gradually diminished from 16.50±2.66 mL/h before splitting to 6.35±1.24 mL/h after splitting. With the exception of TNF-α on postoperative day 2, overall, levels of TNF-α, IL-1, IL-6, and MDA were significantly lower in the NMP group versus CS group for all 5 days postoperatively. Finally, cytochrome C, caspase 3, and NF-κB p65 expressions were all significantly suppressed in the NMP group as compared with the CS group. CONCLUSIONS MP preservation is superior to cold preservation in protecting against liver IRI during RSLT in pigs.

Figure 1

Changes of bile secretion.

Figure 2

TNF-α levels in sera from the blood samples as assessed using ELISA.

Figure 3

IL-1 levels in sera from blood samples as assessed using ELISA.

Figure 4

IL-6 levels in sera from blood samples as assessed using ELISA.

Figure 5

MDA levels in sera from blood samples as assessed using ELISA.

Figure 6

Western blot analysis of relative Cytochrome C expression in liver tissue of the CS and NMP groups as determined after splitting, after reperfusion and after death (* P<0.05). All experiments were repeated three times.

Figure 7

Western blot analysis of relative caspase 3 expression in liver tissue of the CS and NMP groups as determined after splitting, after reperfusion and after death (* P<0.05). All experiments were repeated three times.

Figure 8

Western blot analysis of relative NF-κB p65 expression in liver tissue of the CS and NMP groups as determined after splitting, after reperfusion and after death (* P<0.05) All experiments were repeated three times.