血红素氧化酶-1在大鼠肝移植中保护作用的研究

Study on Protection of Heme Oxygenase-1 in Rat Liver Transplantation

目的探讨改变血红素氧化酶-1(HO-1)的活性对大鼠同种异体原位肝移植的影响。方法用二袖套法建立大鼠同种异体原位肝移植模型,实验分为4组:实验对照组(n=44):供体鼠术前24h腹腔注射生理盐水5ml/kg;氯化血红素组(n=44):供体鼠术前24h腹腔注射氯化血红素100mg/kg,在供肝4℃生理盐水保存期间,经门静脉注入氯化血红素100mg/kg;锌原卟啉组(n=44):供体鼠术前24h腹腔注射锌原卟啉5mg/kg,在供肝4℃生理盐水保存期间,经门静脉注入锌原卟啉5mg/kg;正常大鼠作为正常对照组(n=5)。分别应用RT-PCR及免疫组化技术检测移植肝脏中HO-1mRNA及蛋白表达,并用流式细胞仪分析移植肝脏肝细胞的凋亡率。结果肝移植术后供体肝HO-1表达增强。肝移植术后氯化血红素组HO-1mRNA表达水平高于实验对照组,血清ALT和AST水平低于实验对照组(P<0.05);锌原卟啉组术后肝组织HO-1mRNA表达水平低于实验对照组(P<0.05),血清ALT和AST水平高于实验对照组(P<0.05)。肝移植术后48h肝细胞凋亡率锌原卟啉组最高,氯化血红素组最低,实验对照组介于二者之间,组间差异有统计学意义(P<0.05)。术后7d实验对照组、氯化血红素组及锌原卟啉组3组的生存大鼠分别为7/12、9/12和4/12,组间差异均有统计学意义(P<0.05)。结论肝移植手术过程可促进大鼠肝脏HO-1表达增强;术前采用药物诱导HO-1可降低移植术后肝细胞凋亡率,减轻肝脏损伤程度,提高术后生存率。

Objective To investigate the effect of heine oxygenase 1 （HO-1） activity on rat liver transplantation model. Methods One hundred and thirty-seven rats were divided into 4 groups. Study control group （n=44） ： 24 h before operation, saline 5 ml/kg was infused into peritoneal cavity of donor rats; Heroin group （n= 44） ： heroin 100 mg/kg was infused into peritoneal cavity of donor rats 24 h before operation, and heroin 100 mg/kg was infused into portal vein during the preserve time in 4℃ saline; ZnPP group （n=44） ： ZnPP 5 mg/kg was infused into peri toneal cavity of donor rats 24 h before operation, and ZnPP 5 mg/kg was infused into portal vein during the preserve time in 4 C saline; Normal control group （n=5）： normal rats as normal control group. Expressions of HO-1 protein and mRNA were detected with immunohistoehemistry and RT-PCR technique respectively. The apoptosis rate of hepatocytes was analyzed by flow cytometry. Results Expression of HO-1 mRNA in the liver of heroin group after transplantation was higher than that in study control group obviously, serum ALT and AST levels were lower than those in study control group （P〈0.05） ; HO-1 mRNA expression in ZnPP group liver was lower than that in study control group, serum ALT and AST levels were higher than those in study control group （P〈0.05）. About liver cell apoptosis rate 48 h after liver transplantation, ZnPP group was the highest, hemin group was the mini mum, and there had a significant difference between two groups （P〈0.05）. Seven days after transplantation, the survival ratios of control study group, hemin group and ZnPP group were 7/12, 9/12 and 4/12 in turn, the intergroup differences had statistical significance （P〈0.05）. Conclusion Activity of HO-1 could be induced by the transplant operation. HO-1 increases the survival rate after liver transplantation which was related with reducing apoptotic ratio of hepatocyte and improve hepatic function.