血红素氧合酶-1基因转染对大鼠供肝缺血再灌注损伤的抗凋亡作用

目的 应用转染血红索氧合酶-1(HO-1)基因研究其在大鼠供肝缺血再灌损伤(IRI)中的抗凋亡作用.方法 将转染HO-1基因的腺病毒载体和空载体分别注入供体大鼠腹腔(n=8),36 h后取肝冷保存4 h行大鼠原位肝移植.缺血再灌注6 h检测肝功能、肝细胞凋亡率和肝组织HO-1、bcl-2、hel-xl和Caspase-3的表达.结果 (1)实验组的肝功能明显改善、肝细胞凋亡率显著低于对照组(1.09±0.28)%比(8.30±1.08)%,P<0.01].(2)Western blot检测肝组织HO-1、bel-2和bcl-xl,灰度比值实验组显著高于对照组(HO-1:0.275±0.065比0.035±0.03;bcl-2:0.275±0.025比0.06±0.07;bcl-xl:(0.099±0.041比0.064±0.064,P<0.01),而Caspase-3则显著低于对照组(0.08±0.04比0.21±0.09,P<0.01).结论 HO-1在供肝IRI中通过上调bel-2、bel-xl和下调Caspase-3对供肝有显著的抗凋亡保护作用.

Human heine oxgygenase-1 gene transfer protects rat grafts from IRI by antiapopteotic effect followed by OLT

Objective To investigate the antiapoptopic effect of heme oxgygenase-1 (HO-1) gene transfer in a rat model of IRI followed by orthotopic liver transplantation (OLT). Methods Control and experimental donors (n = 8) were intraperitoneally pretreated with Ad-EGFP and Ad-HO-1 respective-ly 36 h before their livers were harvested. Six h after reperfusion, serum ALT, AST and LDH levels were determined and the apoptotic ratio was analyzed by flow cytometer. The expression of HO-I, bel-2, bcl-xl and Caspase-3 was detected by Western-blot. Results (1) The expression level of HO-1 in the experi- mental group was significantly higher than in the control group (0.275±0.065 vs 0.035±0.03, P <0.01). The liver function index was meliorated and the apoptotic ratio of hepatic cells decreased signifi-cantly (8.30±1.08) % vs (1.09±0.28) %] in the experimental group as compared with those in the control group; (2) The expression of bcl-2 and bcl-xl in the experimental group was increased significant-ly (bcl-2:0.06±0.07 vs 0.275±0.025;bcl-xl:0.064±0.064 vs 0.099±0.041 ,P <0.01) ,and the expression of casepase-3 was weakened obviously (0.21±0.09 vs 0.08±0.04, P < 0.01) as compared with those in the control group. Conclusion HO-1 can alleviate IRI in rat liver by suppressing apoptosis which relates to significantly modulating proapoptotic (Caspase-3) and anfiapoptotic (bcl-2 and bcl-xl) pathways.