缺血预适应对肝脏热缺血再灌注损伤保护作用的机制研究  

The pathogenesis of ischemic preconditioning to warm ischemia reperfusion injury of hepatocytes in rats

作　　者：金涛[1] 胡宇[2] 刘超[3] 

机构地区：[1]天津市南开医院重症医学科,300100 [2]解放军第452医院麻醉科 [3]天津市胸科医院心内科

出　　处：《天津医药》2016年第10期1233-1237,共5页Tianjin Medical Journal

基　　金：2013年中国博士后科学基金资助项目(2013M530880);2015年中国博士后科学基金资助项目(2015M581308)

摘　　要：目的探讨缺血预适应对肝脏热缺血再灌注损伤保护作用的分子机制。方法 90只成年清洁级SD大鼠随机均分为3组,A组为假手术组,B、C组建立SD大鼠肝脏热缺血再灌注损伤模型,并对C组大鼠进行缺血预适应干预。于缺血再灌注后0、2、6、12、24 h采集血标本测定丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST),采用酶联免疫吸附(ELISA)技术检测肝细胞中肿瘤坏死因子(TNF)-α及白细胞介素(IL)-1β水平;采集肝组织标本测定丙二醛(MDA)和超氧化物歧化酶(SOD),流式细胞仪测定肝细胞线粒体膜电位。结果 B、C组大鼠的血清转氨酶、TNF-α、IL-1β及MDA水平均明显高于A组(P<0.05);B、C组大鼠的凝血酶原活动度和胆碱酯酶低于A组(P<0.05);B、C组大鼠肝脏SOD水平明显低于A组;C组大鼠各项指标均优于B组(P<0.05)。缺血再灌注发生后肝细胞线粒体膜电位水平在0 h后即达到最低值,此后呈逐渐上升趋势(P<0.05),并且C组的肝细胞线粒体膜电位水平在各时段均高于B组(P<0.05)。结论缺血预适应对肝细胞热缺血再灌注损伤具有一定的保护作用,缺血预适应可以通过减少炎症因子TNF-α及IL-1β的释放,提高SOD拮抗自由基的活性,减轻线粒体损伤等途径发挥其保护作用。Objective To explore the pathogenesis of ischemic preconditioning to warm ischemia reperfusion injury of hepatocytes in rats.Methods Ninety SD rats were randomly divided into three experimental groups:sham operation group(group A),warm hepatic ischemia/reperfusion group(group B and group C).Group C was given ischemic preconditioning treatment.Serum levels of alanine aminotransferase(ALT) and aspartate aminotransferase(AST) were detected 0 h,2 h,6 h,12 h and 24 h after ischemia reperfusion injury.Levels of TNF-α and IL-1β were tested detected by ELISA.Levels of malondialdehyde(MDA) and superoxide dismutase(SOD) of hepatocytes were detected at the same time points.Mitochondrial membrane potential was examined to assess ischemia reperfusion injury of hepatocytes in rats using chart of intensity of JC-1 in mitochondria.Results The serum levels of ALT,TNF-α,IL-1β,and MDA were significantly higher in hepatic warm ischemia reperfusion group and ischemic preconditioning group than those in sham operation group(P<0.05).Values of prothrombin activity and cholinesterase were significantly lower in liver warm ischemia reperfusion group and ischemic preconditioning group than those of sham operation group(P<0.05).The SOD level of liver was significantly lower in warm ischemia reperfusion group and ischemic preconditioning group than that in sham operation group.The indexes were better in ischemic preconditioning group than those of warm ischemia reperfusion group(P<0.05).The mitochondrial membrane potential level of liver cells reached the lowest value 0 hours after ischemia and reperfusion,and then increased gradually within 24 hours(P<0.05).And the level of mitochondrial membrane potential of liver cells was significantly higher in ischemic preconditioning group than that in warm ischemia reperfusion group(P<0.05).Conclusion Ischemic preconditioning may play a protective role in warm ischemia- reperfusion injury in rats.Ischemic preconditioning may significantly decrease the levels of ALT,AST,TNF-α,IL-1β and MDA,and i

关 键 词：丙氨酸转氨酶 天冬氨酸氨基转移酶类 再灌注损伤 氧化性应激 线粒体 缺血预适应 

分 类 号：R657.3[医药卫生—外科学]