供肝免冲洗灌注的大鼠肝移植实验研究

Experimental study of liver transplantation without graft irrigation or perfusion in rat models

目的探讨供肝免冲洗灌注的大鼠肝移植的可行性。方法将72只健康成年SD大鼠随机分为3组:供体全身肝素化的供肝免冲洗灌注(NFLS组),供体不行全身肝素化的供肝免冲洗灌注(NFL组),供体行全身肝素化的供肝UW液冲洗灌注(对照组)。每组各24只,组内大鼠随机作为供受体进行同种异体大鼠肝移植。分别于移植术后24、72 h获取组织及血清标本送检,检测血清AST、ALT及TB;同时观察各组供肝术后大体观和组织病理学变化。3组AST、TB等肝功能指标比较采用Kruskal-Wallis秩和检验,两两比较采用Wilcoxon检验。结果3组均顺利完成肝移植,术后受体均顺利存活。NFLS组受体术后72 h血清AST、TB水平分别为112(89,181)U/L、0.5(0.4,0.6)μmol/L,明显低于对照组的453(361,692)U/L、1.8(1.5,2.2)μmol/L(Z=-2.56,-2.89;P<0.05)。3组术后24、72 h移植肝灌注良好,移植肝表面均未见明显坏死灶。HE染色显示NFLS组和对照组移植肝肝小叶结构正常,仅有轻微肝脏小叶中心微空泡脂肪变性;NFL组移植肝肝窦内部可观察到微循环血栓形成。结论与传统的器官保存液冲洗灌注相比,免冲洗灌注大鼠肝移植术后肝功能和组织学损伤并无明显加重,供肝免冲洗灌注肝移植可能具有一定可行性,供体全身肝素化是其关键条件。

Objective To investigate the feasibility of liver transplantation without liver graft irrigation or perfusion in rat models.Methods 72 healthy adult SD rats were randomly and evenly divided into three groups:non-flushed donor liver with systemic heparinization(NFLS group),non-flushed donor liver without systemic heparinization(NFL group),and systemic heparinization with UW solution irrigation and perfusion(control group).In each group,the rats were randomly assigned as liver transplantation donors and recipients.The tissue and serum samples were obtained at 24 h and 72 h after liver transplantation for AST,ALT and TB detection.Gross and histopathological changes of liver grafts were observed after liver transplantation.The liver function indexes,such as AST and TB,were compared by Kruskal-Wallis test,and pairwise comparison was performed by Wilcoxon test.Results Liver transplantation was successfully completed in all three groups,and all recipients survived after surgery.In the NFLS group,serum AST and TB levels were 112(89,181)U/L and 0.5(0.4,0.6)μmol/L at postoperative 72 h,significantly lower than 453(361,692)U/L and 1.8(1.5,2.2)μmol/L in the control group(Z=-2.56,-2.89;P<0.05).At postoperative 24 and 72 h,the liver grafts were properly perfused in three groups,and no evident necrosis was found on the surface of liver grafts.HE staining showed that the structure of liver lobules of grafts was normal in the NFLS and control groups,with slight fatty degeneration of microvesicles in the center of liver lobules.In the NFL group,microcirculatory thrombosis could be observed in the sinus of liver grafts.Conclusions Compared with conventional irrigation and perfusion using organ preservation solution,liver function and histological damage are not significantly aggravated after undergoing liver transplantation without irrigation or perfusion in rats,which might bea feasible option in clinical practice.Systemic heparinization of the donor is the key procedure.