转染血红素氧合酶-1基因减轻大鼠脂肪肝移植后的缺血再灌注损伤  

Heme oxygenase-1 gene transfer attenuates rat steatotic liver graft from ischemia/reperfusion injury

作　　者：肖江卫[1] 夏树森[1] 金先庆[2] 王崇树[1] 魏寿江[1] 周彤[1] 王城[1] 李忆秋[1] 

机构地区：[1]川北医学院附属医院普外科,四川南充637000 [2]重庆医科大学附属儿童医院普外科

出　　处：《中华器官移植杂志》2008年第11期652-656,共5页Chinese Journal of Organ Transplantation

基　　金：国家自然科学基金资助项目（30700773）

摘　　要：目的探讨转染血红素氧合酶1（HO-1）基因对大鼠脂肪肝移植后缺血再灌注损伤的影响。方法利用分子生物学方法构建携带有HO-1基因的重组腺病毒（AdS—HO-1），于供肝切取前48h经阴茎背静脉将Ad5-HO-1注入供者体内，供肝置于4℃HTK液保存2h，然后移植。对照组接受正常肝移植；轻度对照组接受轻度脂肪肝移植；轻度实验组接受注射Ad5-HO-1的轻度脂肪肝移植；重度对照组接受重度脂肪肝移植；重度实验组接受注射Ad5-H＆I的重度脂肪肝移植。术后观察各组大鼠的存活率及肝功能；观察移植肝组织的病理变化；测定移植肝组织中HO-1的活性以及HO-1、Bcl-2、锌指蛋白A20、凋亡蛋白酶3（Caspase-3）的表达。结果与重度对照组相比较，重度实验组的丙氨酸转氨酶水平显著下降（P〈O．05）。轻度实验组和重度实验组的HO-1活性分别高于各自的对照组（P〈0．05）。重度实验组移植肝组织中HO-1、Bcl—2及锌指蛋白A20的表达水平明显高于重度对照组（P〈O．05），而Caspase-3的表达是降低的（P〈0．05）。重度实验组的1、7、21d存活率分别为66．7％（4／6）、50％（3／6）和50％（3／6），而重度对照组的1、7、21d存活率分别为33．3％（2／6）、16．7％（1／6）和16．7％（1／6），二者比较，差异有统计学意义（P〈0．05）。结论转染血HO-1基因可减轻大鼠脂肪肝移植后的缺血再灌注损伤。Objective To understand the role of heme oxygenase-1 （HO-1） during ischemia reperfusion injury （IRI） after steatotic liver transplantation, we up-regulated the HO-1 expression of donors by gene transfection before transp]antatior/and studied the effects and mechanisms of HO-1 during IRI. Methods Recombinant adenoviral vector encoding HO-1 gene （Ad5-HO-1） was constructed by using molecular biology method, and administered to donors yia penile vein at 48 h before transplantation. Livers of the donors were harvested and stored for 2 h at 4 ℃ in HTK solution before they were implanted into the recipients. Five groups were set up.. control group （normal liver graft to be implanted）, the mild steatosis liver control group （mild fatty liver graft to be implanted）, the mild steatosis with Ad5-HO-1 pretreatment group （rats to be pretreated with AdS-HO-1 and mild fatty liver graft to be implanted）, the severe steatosis liver control group （severe fatty liver graft to be implanted） and the severe steatosis with AdS-HO-1 pretreatment group （rats to be pretreated with Ad5-HO-1 and severe fatty liver graft to be implanted）. The survival rate of rats and liver graft pathological changes were observed. The levels of serum ALT, AST, TB, activity of HO-1, and the expression levels of HO-1, Bcl-2, zinc finger protein A2O, Caspase-3 were assayed. Results Compared with the severe steatosis liver control group, levels of serum ALT in the severe steatosis with Ad5-HO-1 pretreatment group were significantly reduced （P%0. 05）; Activity of HO-1 in the severe steatosis with Ad5-HO-1 pretreatment group and the mild steatosis with Ad5 HO-1 pretreatment group were significantly increased （P 〈 0. 05）. Western blot and RT-PCR assays revealed that the expression of HO-1, Bcl-2 and A20 in the severe steatosis with AdS-HO-1 pretreatment group was stronger than in the severe steatosis liver control group （P〈0. 05）, but that of Caspase-3 weaker. The survival rate in the severe steatosis with Ad5-HO-1

关 键 词：血红素氧化酶 肝移植 再灌注损伤 动物 基因修饰 大鼠 

分 类 号：R686[医药卫生—骨科学]