Establishment And Protective Mechanism Of Hypothermic Oxygenated Machine Perfusion System In Mouse Liver

Objective Due to insufficient source of organs,donation after circulatory death(DCD)liver and other poor quality livers were used in clinical applications.To compensate the deficiency of cold storage(CS)in repairing liver quality,hypothermic oxygenated machine perfusion(HOPE)technology was gradually being used.However,because of unsuitable research models for further research,the molecular mechanism of HOPE was still unclear.To this end,we intend to establish a mouse liver HOPE model,based on the exploration of appropriate perfusion parameters,the model is used to clarify the protective mechanism of HOPE on DCD donor liver.Methods Through comparing the effects of different perfusion flow on normal livers or warm ischemia 30 minutes livers,we verified the stability and effectiveness of mouse liver HOPE system,and explored the appropriate perfusion flow.The role of autophagy in HOPE was demonstrated by using a normothermic reperfusion system without blood in vitro.Through utilizing a normothermic reperfusion system with 25% blood in vitro and the gene knockout mouse,we analyzed the function of P-selectin HOPE's protective effect on DCD donor livers.Results When the flow of HOPE was not less than 0.3 ml/min·g,the system could achieve complete perfusion in the normal liver.Under the condition of flow between 0.1 ml/min·g and 0.5 ml/min·g,HOPE could exert a significant protecting role in DCD livers.Compared with CS,the DCD iver undergoing HOPE could activate the production of autophagosome and the expression of autophagy protein.However,the application of autophagy inhibitor could eliminate the protective effect of HOPE.In terms of inflammatory response,P-selectin knockout could attenuate DCD injury,which had similar protective effects of wild-type mouse HOPE.In addition,comparing the effect of HOPE and CS on DCD livers in P-selectin knockout mice,P-selectin knockout mouse HOPE could also alleviate DNA and oxidative stress injury except for anti-inflammatory effect.Conclusion We have established a stable mouse liver HOPE system successfully.We also clarified that HOPE could protect DCD livers by activating autophagy and P-selectin-dependent and-independent pathways.