Hypothermic Machine Perfusion(HMP) attenuates liver inflammation response after reperfusion through upregulating expression of KLF2

Abstract

Objective The aim of this research was to investigate the expression of KLF2 and nuclear factor- kappa B P65(NF-κB P65) in hypothermic machine perfusion and its role in liver inflammatory response after rat liver transplantation from non-heart-beating donor. Methods Twelve rats were randomly divided into two groups: HMP group(n=6) and cold storage (CS) group(n=6). With 30min of hepatic pedicle occlusion, the rats were sacrificed and liver were obtained. HMP group experienced HMP and CS group experienced CS for 3h, respectively. Using the Isolated perfused rat liver model (IPRL) to mimick the reperfusion after transplantation. During normothermic reperfusion, intrahepatic resistance (IR) was calculated, the activities of hepatic enzyme in perfusate and oxygen consumption were monitored and production of bile was evaluated. After normothermic reperfusion, livers were obtained for histological study, the expression of Kruppel-like Factor 2(KLF2) and NF-κBP65 were detected by Western Blot and the expression of TNF-α and IL-1β in liver were detected by ELISA. Results 1. After 0min, 60min, 120min of reperfusion, levels of ALT、AST、LDH in HMP group were significant lower than these in CS group (P 0.05). 2. In HMP group, hepatic sinusoids were somewhat dilated and only a very few hepatic cell edema, while CS group showed a serious liver injury including large hepatic cell edema, cytoplasmic loose, congestion and inflammation cells in the hepatic sinusoidal. 3. The expression of KLF2 in HMP group were significantly higher than CS group(P<0.05), while the expression of NF-κBP65、TNF-αand IL-1βin HMP group were significantly lower than CS group (P<0.05). Conclusion HMP could decrease liver inflammation response after liver reperfusion of non-heart-beating donor through upregulating KLF2 and inhibiting NF-κB p65 expression. Key words: Rats; Liver; Machine perfusion; Cold preservation; Kruppel like factor 2