The Study On The Application Of Hydrogen-enriched Organ Preservation Solution In Cold Storage And Reperfusion Of Rat Graft Liver

Background:The cold storage and reperfusion injury are the most common reasons for the occurrence of primary liver graft nonfunction and delayed graft function restoration after liver transplantation.The injury mechanism is believed to be redox imbalance and secondary inflammatory reaction.Several studies have proven that hydrogen gas?H₂?is able to alleviate oxidative stress injury and inflammatory reaction.However,there is few hydrogen-related researches in liver transplantation field currently and therefore further studies on its efficiency and mechanism should be performed.Aim:The study aim to verify the characteristics of the injury in different stages throughout hepatic ischemia-reperfusion.Anovelmodified isolated perfusion of rat liver?MIPRL?model is also to be established to evaluate the physiological and biochemical changes of graft liver.The protective effect and mechanism of hydrogen gas on various phases during cold storage and reperfusion of graft liver is also to be explored.Differences of protective effect using hydrogen between elderly and young liver graft are planed to be compared.The application profile for hydrogen-enriched cold storage and reperfusion with high clinical translational potential is planed to be established.Methods:The study firstly used rat liver ischemia-reperfusion model and MIPRL model to explore characteristics of the injury in different phases.Hydrogen-enriched Celsior?preservation solution and Krebs-Henseleit perfusate in high concentration was produced using hydrogen generation agent.The most favorable reaction phases were explored by using hydrogen-enriched preservation solution and perfusate jointly or alone in different phases.Hydrogen gas was also applied on elderly graft liver to verify the existing of protective effect and the difference between elderly and young graft liver.MIPRL was used to evaluate liver condition.Level of ALT,AST,PVP/LW,bile production in perfusate and HPO in liver tissue were measured.After perfusion,levle of oxidative stress-related index and inflammatory reaction were determined.Both of pathological injury and apoptotic situation were examined.Results:In the rat liver ischemia-reperfusion model,the level ofALT,AST and MDAin serum at4 h after reperfusion were higher than those during ischemia phase,as well as SOD declined.In MIPRL model,level ofALT,AST,HPO,MDAand PVP/LW in graft liver perfusate increased and SOD declined,with severe pathological injury,after cold storage and reperfusion.When hydrogen-enriched preservation solution and perfusate were used jointly,ALT,AST,MDA,bile production and TNF-? were apparently lower than those in groups using hydrogen-enriched solution or perfusate alone,with increased SOD and GSH/GSSG level.Cellular edema,apoptosis and structure damage decreased.The SOD level in elderly liver was lower than young liver.Comparing with group without hydrogen,level ofALT,AST,HPO and GSSG decreased and SOD and GSH/GSSG increased after using hydrogen gas.Cellular edema,apoptosis and structure damage decreased.Conclusion:Application of hydrogen-enriched preservation solution and perfusate jointly could alleviate cold storage and ischemia-reperfusion injury of young and elderly graft liver effectively.