常温机械灌注减轻心死亡猪供肝缺血再灌注损伤

Normothermic machine perfusion reduces ischemic reperfusion injury of DCD liver grafts in pig liver transplantation

目的 探讨常温机械灌注系统（NMP）减轻心死亡猪供肝缺血再灌注损伤及改善肝移植预后的作用.方法 选用健康巴马小型猪24只,采用随机数字表法将实验动物分为两组：（1）NMP组：6对,无心跳供猪（钳夹心脏基底部主动脉后静脉推注质量分数10％氯化钾30ml）热缺血40 min后切取供肝,连接NMP保存装置常温保存6h后,以4℃的乳酸林格氏液冲洗供肝,行无转流原位肝移植术;（2）UW液组：6对,无心跳供猪经热缺血40 min后常规原位冷灌注后切取供肝,0～4℃UW液保存6h,行无转流原位肝移植术.检测灌注过程中NMP组供肝胆汁分泌情况、供肝组织病理学改变及灌注液生化指标变化,比较术中及术后5d内两组受猪的存活率以及血清生化指标、肝组织病理学变化.结果 NMP组供肝在体外灌注保存过程中,灌注保存液中的丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）及乳酸脱氢酶（LDH）水平等指标在保存的开始出现一定水平的升高,之后维持在较稳定的水平,胆汁分泌量为（10.3±3.61）ml/h,供肝保存6h后组织结构基本正常.NMP组和UW液组术后5d存活率分别为100％和0,两组比较,差异有统计学意义（P＜0.05）.再灌注后NMP组血清生化指标ALT、AST、LDH和乳酸水平低于UW液组,两组比较,差异均有统计学意义（P＜0.05）.肝脏常规病理检查提示,NMP组肝组织细胞损伤程度较UW液组明显减轻.结论 常温机械灌注保存可明显减轻心死亡猪供肝的缺血再灌注损伤,可在体外对供肝进行修复,改善肝移植预后.

Objective To investigate the effect of normothermic machine perfusion （NMP） in alleviating the ischemia reperfusion injury of DCD grafts.Methods Twenty-four Bama miniature pigs were randomly divided into two groups.The donor livers were harvested after 40-min cardiac arrest and preserved for 6 h by NMP preservation in the group A,or with cold UW solution in the group B,respectively.Then,liver transplantation was performed in both groups.Choleresis and the perfusate biochemical markers in the group A were detected during NMP.The recipient survival rate,the serum biochemical markers as well as the histopathological changes of the donor livers were compared between these two groups.Results The levels of ALT,AST and LDH in the perfusate were slightly elevated at the beginning of NMP and maintained at a stable level thereafter.The average choleresis rate was 10.3 ＆#177; 3.61 mL/h.The histological structures of the donor livers were normal after 6-h NMP preservation.The survival rate of the recipient pigs was 100% and 0 in the group A and group B,respectively.After reperfusion,the levels of serum ALT,AST,LDH and lactic acid in the group A were significantly lower than those in the group B （P 〈 0.05）.The pathological examination demonstrated that the hepatocyte injury in group A was slighter than that in group B after 6-h preservation and reperfusion.Conclusion NMP storage could significantly alleviate the ischemia reperfusion injury of pig DCD livers and improve the prognosis of liver transplantation.