肝缺血再灌注后HO-1mRNA的表达及其与细胞凋亡关系的临床研究

Clinical research of the expression of HO-1 mRNA in hepatic ischemic reperfusion injury and the relationship with apoptosis

目的通过检测肝组织在缺血再灌注前后血红素氧合酶-1(HO-1)mRNA的表达及细胞凋亡情况,探讨肝组织在缺血再灌注时HO-1mRNA与细胞凋亡之间的关系。为临床选用有效的预处理方法、抑制细胞凋亡、减轻肝脏缺血再灌注损伤提供理论基础。方法肝切手术中,分别在肝门阻断前、肝门阻断开放同时、手术结束关腹前取切缘旁肝组织10g。逆转录聚合酶链反应(RT-PCR)法检测HO-1mRNA表达。脱氧核糖核苷酸末端转移酶介导的缺口末端标记技术法测定肝细胞凋亡。结果肝组织HO-1mRNA的表达,在肝门阻断前为(0.2313±0.0575),肝门开放时为(0.8168±0.1972),关腹前为(1.2447±0.4403)。这三个组间的差异有显著性(P<0.05)。肝门阻断前HO-1mRNA的表达比肝门开放时少,关腹前HO-1mRNA的表达要比肝门开放时多。肝门阻断前的凋亡指数为(9.590±0.959)‰,肝门开放时为(5.800±0.939)‰,关腹前为(4.690±1.738)‰,这三个时间点的差异有显著性(P<0.05)。肝门阻断前的肝细胞凋亡指数高于肝门开放时和关腹前;而肝门开放时肝细胞凋亡指数高于关腹前。相关性分析:在肝门阻断前、肝门阻断开放同时、手术结束关腹前,肝组织HO-1mRNA的表达与肝细胞凋亡呈高度负相关。结论HO-1作为细胞和组织应激过程中的关键因子,在肝缺血再灌注损伤过程,HO-1可能通过抑制细胞凋亡,减少缺血再灌注时的肝细胞损伤。

[ Objectives ] To detect the apoptosis and the expression of HO-1 mRNA in hepatic tissue during the course of ischemic reperfusion, then discuss the response of hepatic tissue to ischemic reperfusion and the relationship between HO-1 mRNA and apoptosis. To supply the theory of effective pretreatment and lessening hepatic ischemic reperfusion injury. [ Methods] Hepatic tissue （10 gram） was taken around incisal margin before the blockage of porta hepatis, the open of porta hepatis and before the closure of abdominal cavity respectively. The expression of HO-1 mRNA was determined by reverse transcription polymerase chain reaction （RT-PCR）. The hepatoeellular apoptosis was determined by terminal-deoynucleotidle transferase mediated d-UTP nick end labeling method. [ Resuits] The expression of HO-1mRNA in the pre-blockage of porta hepatis group was （0.2313 ± 0.0575）, the open of porta hepatis group was （0.8168 ±0.1972）, the pre-closure of abdominal cavity group was （1.2447 ±0.4403）. Differences among these three groups had distinguished statistical significance （P 〈0.05）. Compared with the open ofporta hepatis group, the expression of HO-1mRNA declined markedly in the pre-blockage of porta hepatis group. Compared with the open of porta hepatis group,the expression of HO-1mRNA increased markedly in the pre-closure of abdominal cavity group. Hepatocyte Apoptosis Index in the pre-blockage of porta hepatic group was （9.590 ±0.959）‰, the open of porta hepatis group was （5.800 ±0.939）‰, the pre-closure of abdominal cavity group was （4.690 ±1.738）‰. Differences among these three groups had distinguished statistic significance（P 〈0.05）. Compared with the pre-blockage of porta hepatis group, Apoptosis Index declined markedly in the open of porta hepatis group. Compared with the open of porta hepatis group, Apoptosis Index declined markedly in the pre-closure of abdominal cavity group. In the pre-blockage of porta hepatis group, open of porta hepatis group and the pre-closure of abdominal cavity group,the expression of HO-1mRNA has a high negative correlation with hepatocyte apoptosis in hepatic tissue.[Conclusion] There were inverse correlation between HO-1 mRNA and the AI in the pre-blockage of porta hepatis group, open of porta hepatis group and the pre-closure of abdominal cavity group. It suggests that HO-1 has an effect on suppressing apoptosis.