| Objective To build NMP(Normothermic machine perfusion,NMP)device for fatty donor liver from pig,and evaluate its effect of preservation and lipid lowering;Preliminarily exploring the intraoperative and postoperative early effect of NMP for fatty donor liver from pig to orthotopic liver transplantation.Methods 1.Building 8 models of Bama minipig with mild fatty liver by feeding with tetracycline + high fat + alcohol for 8 weeks,then randomly divided into two groups after circulatory death.NMP group(n=4)Put the donor liver into the 37 ℃constant temperature NMP device quickly,then perfused portal vein and artery by the perfusate which was diluted with minipig blood for 10h;University of Wisconsin(UW)solution static cold storage group(n = 4),perfused abdominal aorta and portal vein by 1500 ml 4 ℃hypertonic citrate(HCA)organ preservation solutionl and 1000 ml 4 UW solution,℃rapid cooling with ice residue and then preserved by 4 ℃UW solution for 10 h.In NMP group,we observed the changes of hepatic artery and portal vein pressure and flow rate,blood gas analysis,blood glucose correction to maintain the stability of NMP device,acid-base electrolyte,oxygen partial pressure and energy metabolism balance during perfusion;The changes of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in perfusion and the changes of the concentration of triglyceride(TG)and cholesterol(CHO)in the liver tissue homogenate were observed over time;And recorded the bile yield per hour.In SCS group,we observed the levels of triglyceride(TG)and cholesterol(CHO)in liver tissue homogenate before and after perfusion.Both groups had been taken histopathological examination before and after perfusion.Evaluated NMP device’s effect of preservation and lipid lowering.2.Selecting 8 healthy male Bama minipigs as recipients which had same weight、months of age with donor,randomly divided into NMP group(n=4)、 SCS group(n=4).After resection of autologous liver,orthotopic liver transplantation began,NMP group received fatty donor liver which had been normothermic machine perfusion for 12 h,and SCS group took fatty donor liver which had been cold storageby UW solution for 10 h,and took down the basic informations;The liver function indexes,survival rate,mortality and complications were compared between the two groups at the time of opening the artery and postoperative 0h to 120h;The experimental animals were sacrificed at 5 days after operation,and the histopathological data of the transplanted liver were compared.To determine the fatty donor liver experienced normothermic machine perfusion in orthotopic liver transplantation if have advantages than the traditional static cold preservation by UW solution or not.Results 1.NMP group in the process of normothermic machine perfusion,portal vein、hepatic artery pressure and blood flow,temperature control are stable,the p H value(PH),oxygen partial pressure(PO2),electrolyte and blood glucose of the perfusate were kept within the design range.After perfusion,perfusate’s levels of ALT and AST were increased,and decreased slowly at 2h,compared with 0h AST(P = 0.159)and ALT(P = 0.103),10 h perfusion were not statistically significant(P> 0.05);Bile yield after perfusion 0-4h gradually increased,4-10 h slowly reduced.With the perfusion time prolonged,NMP group weight per unit liver tissue homogenate CHO and the mean concentration of TG was lower than that of perfusion 0 h,there was statistical significant difference between perfusion 6h CHO and perfusion 0h(P=0.019)、 perfusion 8h CHO and perfusion 0h.There was no significant difference in weight per unit liver tissue homogenate CHO and TG between the NMP group and the SCS group(P> 0.05).The difference of weight per unit liver tissue homogenate CHO(P = 0.034)and TG(P = 0.046)between the NMP group and the SCS group was statistically significant.After 10 h normothermic machine perfusion or static cold preservation,the two groups of liver routine pathological examination showed that the liver structure was clear,the shape and liver lobule structure were normal;In SCS group the degree of liver cell edema and steatosis,hepatic sinus dilation and congestion were heavier.There were no Significant difference(P> 0.05)in the weight、donor liver mass、 surgical time,anhepatic time,portal vein opening to hepatic artery opening time,intraoperative infusion volume of two group recipient pig.The intraoperative blood loss of NMP group was significantly less than that of SCS group,and the urine volume after opening was significantly higher than that of SCSgroup.0h after liver transplantation,8 recipients all survived,the success rate was 100%;Within 120 hours after surgery,NMP 4 cases all survived,the survival rate was 100%;1 case in SCS group died of 13 h after operation,the survival rate was 75%.In the early stage of postoperation,the levels of ALT and AST in two groups increased differently,SCS group increased more obviously and decreased slowly.ALT and AST were significantly higher in the SCS group than in the NMP group at 1 day to 5 days after operation.In NMP group,TB increased to 2 d after surgery and then began to decline,SCS group began to reduce at the third day after operation;After 2days,NMP group was significantly lower than the SCS group(P <0.05).There was no significant difference between the two groups after 4 days of ALP and GGT(P>0.05),after 96 hours,the NMP group was significantly lower than the SCS group(P>0.05).The survived recipient pigs were sacrificed 5 days after operation.Histopathological examination of transplanted liver showed that the injury degree of hepatic cell in NMP group was lighter than that in SCS group.Conclusion 1.The NMP device in this study run stably,after 10 h perfusion,the lipid content in fatty donor liver from pig DCD decreased,liver function was basically stable,the injury degree of hepatic cell in it was lighter than that in cold preservation,suggested the device have the effect of preservation and lipid lowering.2.In orthotopic liver transplantation,10 h of NMP could relive the reperfusion injury of fatty donor liver from pig DCD to a certain extent at the time of bloodstream opening,promoted the recovery of liver function at the early stage of postoperation,and reduced the primary risk of graft failure. |
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