改良大鼠离体肝脏灌注模型在供肝冷保存中的应用研究  

Establishing A Modified Isolated Perfused Rat Liver Model for Applied in Study of Cold Storage of Liver Grafts

作　　者：施少军 郑建新 秦天 李小康 薛峰[1] 夏强[1] 

机构地区：[1]上海交通大学医学院附属仁济医院肝脏外科,200127 [2]日本国立儿童成长与发育研究所

出　　处：《胃肠病学》2017年第9期517-522,共6页Chinese Journal of Gastroenterology

基　　金：国家自然科学基金(81470847;81670602);上海交通大学医学院转化医学协同创新中心项目(TM201511)

摘　　要：背景:供肝冷保存和缺血再灌注损伤是影响肝移植患者预后的关键因素,建立稳定的冷保存再灌注动物模型是开展相关研究的基础。目的:建立适用于综合评价供肝质量的改良大鼠离体肝脏灌注模型,为供肝冷保存研究提供合适的离体再灌注模型。方法:12只Sprague-Dawley大鼠随机分为两组,对照组正常获取供肝后进行离体灌注90 min,实验组供肝经30 min热缺血后获取,冷保存24 h后再行离体灌注。离体灌注期间监测灌注液转氨酶水平、电解质浓度和pH值;以压力感受器监测供肝门静脉压力;以超微自由基探针监测肝组织过氧化氢(HPO)水平。灌注结束后记录胆汁流出量,检测供肝组织丙二醛(MDA)、超氧化物歧化酶(Cu/Zn SOD)水平及其组织病理学改变和肝细胞凋亡情况。结果:与对照组相比,实验组供肝AST、ALT水平、门静脉压力和HPO水平明显增高,胆汁流出量显著减少,肝组织Cu/Zn SOD水平显著降低。对照组供肝组织病理学损伤和肝细胞凋亡相对较轻。结论:经热缺血和冷保存后,供肝的肝功能和抗氧化能力均明显降低。本研究构建的改良大鼠离体肝脏灌注模型可有效监控和评估大鼠供肝冷保存再灌注损伤情况。Background： Cold storage and ischemia-reperfusion injury of liver grafts are critical for the prognosis of patients undergoing liver transplantation. Establishing a stable cold storage and reperfusion animal model is a fundamental measure for related studies. Aims： To establish a modified isolated perfused rat liver model for comprehensive assessment of liver grafts in studies on preservation of liver grafts before transplantation. Methods： Twelve Sprague-Dawley rats were randomly allocated into two groups. Livers of rats in control group were retrieved and perfused immediately for 90 minutes without preservation. In experimental group,liver grafts underwent a 30-minute warm ischemia followed by 24-hour cold storage before ex vivo perfusion. The perfusate was collected dynamically for monitoring the levels of transaminases,electrolytes and pH value; the portal vein pressure of liver grafts was measured by pressure sensor,and the hepatic hydrogen peroxide（ HPO） level was assessed by microprobe for free radicals. The bile production was recorded after the ex vivo perfusion;meanwhile,the levels of malondialdehyde（ MDA） and superoxide dismutase（ Cu/Zn SOD）,the histopathological changes and apoptosis of hepatocytes of liver grafts were examined. Results： Compared with the control group,the levels of AST and ALT,the portal vein pressure and the HPO level of liver grafts in experimental group were obviously increased throughout the perfusion. Furthermore,the bile production and level of Cu/Zn SOD of liver grafts in experimental group were significantly decreased. The histopathological injury and hepatocytes apoptosis of liver grafts were milder in control group. Conclusions： The liver function and antioxidant effect were reduced in warm ischemic and cold preserved liver grafts. The modified isolated perfused rat liver model established in this study is useful for monitoring and evaluation of the cold storage and reperfusion injury in liver grafts.

关 键 词：肝移植 器官保存 离体灌注 再灌注损伤 大鼠 Sprague-Dawley 

分 类 号：R657.3[医药卫生—外科学]