CAJ | 학술논문

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(Ad5-HO-1),于供肝切取前48 h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2 h,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-HO-1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＜0.05).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＜0.05),而Caspase-3的表达是降低的(P＜0.05).重度实验组的1、7、21 d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、21 d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.
목적 탐토전염혈홍소양합매-1(HO-1)기인대대서지방간이식후결혈재관주손상적영향.방법 이용분자생물학방법구건휴대유HO-1기인적중조선병독(Ad5-HO-1),우공간절취전48 h경음경배정맥장Ad5-HO-1주입공자체내,공간치우4℃HTK액보존2 h,연후이식.대조조접수정상간이식;경도대조조접수경도지방간이식;경도실험조접수주사Ad5～HO-1적경도지방간이식;중도대조조접수중도지방간이식;중도실험조접수주사Ad5-HO-1적중도지방간이식.술후관찰각조대서적존활솔급간공능;관찰이식간조직적병리변화;측정이식간조직중HO-1적활성이급HO-1、Bcl-2、자지단백A20、조망단백매-3(Caspase-3)적표체.결과 여중도대조조상비교,중도실험조적병안산전안매수평현저하강(P＜0.05).경도실험조화중도실험조적HO-1활성분별고우각자적대조조(P＜0.05).중도실험조이식간조직중HO-1、Bcl-2급자지단백A20적표체수평명현고우중도대조조(P＜0.05),이Caspase-3적표체시강저적(P＜0.05).중도실험조적1、7、21 d존활솔분별위66.7%(4/6)、50%(3/6)화50%(3/6),이중도대조조적1、7、21 d존활솔분별위33.3%(2/6)、16.7%(1/6)화16.7%(1/6),이자비교,차이유통계학의의(P＜0.05).결론 전염혈HO-1기인가감경대서지방간이식후적결혈재관주손상.
Objective To understand the role of heme oxygenase-1(HO-1)during ischemia reperfusion injury(IRI)after steatotie liver transplantation,we up-regulated the HO-1 expression of donors by gene transfection before transplantatiori and studied the effects and meehanisms 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 fatry liver graft to be implanted),the mild steatosis with Ad5-HO-1 pretreatment group(rats to be pretreated with Ad5-HO-1 and mild fatty Iiver graft to be implanted).the severe steatosis Iiver controI group(severe fatty liver graft to be implanted)and the severe steatosis with Ad5-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 Iiver 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 A20,caspase-3 were assayed.Results Compared with the severe steatosis liver eontrol 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 Ad5-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 pretreatment group at 1 st,7th,and 21 st day was respectively 66.7%(4/6),50%(3/6),50%(3/6),and that in the severe steatosis liver control group was respectively 33.3%(2/6),16.7%(1/6),16.7%(1/6)(P＜0.05).Conclusion HO-1 gene transfer could attenuate rat steatotic livers from IRI after rat liver transplantation.