血红素氧合酶-1对大鼠肝移植缺血再灌注损伤后缺氧诱导因子-1α和血管内皮生长因子表达的影响  

Ischemia-reperfusion injury; Liver transplantation; Heine oxygenase-1 ; RevascularizationEffect of heme oxygenase-1 on expressions of hypoxia inducing factor 1 alpha and vascular endothelial growth factor 1 alpha after orthotopic liver transplantation ischemia-reperfusion injury in rats

作　　者：张治清 展希[1] 黄汉飞[1] 段键[1] 张玉君[2] 王昆华[1] 曾仲[1] 

机构地区：[1]昆明医科大学第一附属医院器官移植中心,650032 [2]第三军医大学西南医院全军肝胆外科研究所,重庆400038

出　　处：《中华消化外科杂志》2017年第9期955-962,共8页Chinese Journal of Digestive Surgery

基　　金：国家自然科学基金项目（81360079）;云南省肝、肾移植创新团队（2009C1009）

摘　　要：目的探讨血红素氧合酶-1（HO-1）对大鼠肝移植缺血再灌注损伤后肝脏组织中缺氧诱导因子-1α（HIF-1α）、VEGF的表达和肝内血管再生的影响。方法采用实验研究方法。240只大鼠按随机数字表法，分为3组，每组80只，空病毒组大鼠转染空病毒，诱导组大鼠转染HO-1过表达腺病毒，抑制组大鼠转染HO-1RNAi腺病毒。大鼠进行1：1配对，选取体质量较小者作为供体，体质量较大者作为受体，参照“二袖套”法大鼠肝移植模型行肝移植。供、受体大鼠均于手术前24h经尾静脉注射腺病毒。（1）术前腺病毒转染效率检测：注射腺病毒后12、24h，Westernblot检测3组大鼠肝脏组织中HO-1的表达量。（2）肝移植术后受体大鼠肝功能检测：肝移植术后1、3、7、14d检测3组受体大鼠肝功能指标（ALT、AST、ALP、GGT）。（3）肝移植术后受体大鼠肝组织病理形态学和损伤评分：术后1d和14d取受体大鼠肝脏组织制备成石蜡切片，HE染色观察，并采用Suzuki损伤评分标准评价。（4）Westernblot检测受体大鼠肝脏组织中HIF-1α、VEGF、HO-1相对蛋白表达量。（5）免疫荧光染色检测3组受体大鼠肝移植术后14d肝脏组织中血管性血友病因子（vWF）。计数小血管数量。正态分布的计量资料以孟±s表示，两组间比较采用独立样本的t检验，多组间比较采用单因素方差分析。两两比较采用LSD检验。结果（1）术前腺病毒转染效率检测：空病毒组、诱导组和抑制组大鼠术前注射腺病毒12h时肝脏组织中HO-1相对表达量分别为1．08±0．16、1．18±0．21、0．87±0．26，24h时分别为1．08±0．26、1．39±0．19、0．57±0．12，3组不同时间点比较，差异均有统计学意义（F=4．232，36．513，P〈O．05）。（2）肝移植术后受体大鼠肝功能情况：肝移植术后3d空病毒组、诱导组和抑制组大鼠ALT分别为（504±67）U／L、（438±47）U／L�Objective To explore the effect of heine oxygenase-1 （HO-1） on expressions of hypoxia inducing factor 1 alpha （HIF-1α） and vascular endothelial growth factor （VEGF）and regeneration of hepatic vascular plexus after orthotopic liver transplantation ischemia-reperfusion injury in rats. Methods The experimental study was conducted. According to the random number table, 240 SD rats were divided into the 3 groups, 80 rats in each group. Empty virus group： rats were transfected with the empty virus. Induced group： rats were transfected with HO-1 overexpression adenovirus. Inhibited group： rats were transfected with HO-1 RNAi adenovirus. Rats were made pairs （ 1 ： 1 ） and established rat liver transplantation model according to ＂two cuffs method＂. Rats with less weight and with heavier weight were respectively chosen as donor rats and recipient rats, and then recieved tail intravenous injection of adenovirus at 24 hours before operation. （1） Detection of transfection efficiency of adenovirus before operation： HO-1 expression of liver tissue of rats in each group was detected by Western blot at 12 and 24 hours after injection of adenovirus. （2） Liver function test of recipient rats after liver transplantation ： liver functions of recipient rats [ alanine transaminase （ALT） , aspartate transaminase （ AST）, alkaline phosphatase （ALP）, gamma-glutamyl transferase （GGT） ] were detected at 1, 3, 7 and 14 days postoperatively. （3） Pathological histology of liver tissue and injury scores of recipient rats in the 3 groups after liver transplantation： paraffin sections of recipient rats in the 3 groups at postoperative 1 and 14 days were stained by HE staining and observed by light microscope, and were evaluated by Suzuki damage score standard. （4） Relative expressions of HIF-1α, VEGF and HO-1 in liver tissue of recipient rats were detected by Western blot. （5） Von Willebrand factor （vWF） in liver tissue of recipient rats at 14 days postoperatively was dete

关 键 词：缺血再灌注损伤 肝移植 血红素氧合酶-1 血管再生 

分 类 号：R657.3[医药卫生—外科学]