缺血预处理对大鼠肝脏缺血再灌注损伤的保护作用及对缺氧诱导因子1α蛋白表达的影响  

Protective effects of ischemic preconditioning on the injury of hepatic ischemia reperfusion in rats and its influence on the expression of hypoxia-inducible factor 1α protein

作　　者：张彬[1] 梁静[2] 王坤[3] 孙培胜[1] 朱绍辉[1] 

机构地区：[1]新乡医学院第一附属医院普通外一科,河南卫辉453100 [2]新乡医学院第三附属医院皮肤科,河南新乡453003 [3]新乡市第一人民医院普通外科,河南新乡453000

出　　处：《新乡医学院学报》2014年第6期409-413,共5页Journal of Xinxiang Medical University

基　　金：河南省教育厅自然科学项目(编号:2011a32001);河南省教育厅自然科学项目(编号:2012b320019);河南省教育厅自然科学项目(编号:14A320075);河南省卫生厅科技攻关项目(编号:201203071)

摘　　要：目的探讨缺血预处理(IP)对肝脏缺血再灌注(IR)损伤的保护作用及对缺氧诱导因子1α(HIF-1α)蛋白表达的影响。方法将120只Sprague-Dawley大鼠分为假手术(SO)组、IR组和IP组,每组40只。SO组术中只牵拉分离肝十二指肠韧带,操作完成后关腹;IR组阻断十二指肠韧带,造成缺血30 min后解除阻断;IP组在阻断肝十二指肠韧带前30 min进行持续5 min缺血及5 min再灌注。IR和IP组分别于再灌注2、6、12和24 h时间点各处死10只动物,取肝脏标本;SO组于术后2 h取肝组织标本。检测血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、肿瘤坏死因子-α(TNF-α)、白细胞介素-10(IL-10)水平,免疫组织化学法测HIF-1α蛋白表达水平,用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记测定法(TUNEL)测定肝细胞凋亡,并计算细胞凋亡指数(AI)。结果再灌注后,IR组及IP组大鼠各时点的ALT、AST、TNF-α、IL-10与SO组相比均升高,差异均有统计学意义(P<0.05)。IP组大鼠各时间点TNF-α明显低于IR组,IL-10明显高于IR组(P<0.05)。IP组大鼠ALT和AST在2、6、12 h低于IR组,差异均有统计学意义(P<0.05)。IR组和IP组大鼠细胞AI较SO组增加,差异有统计学意义(P<0.05);IP组大鼠各时间点AI低于IR组,差异均有统计学意义(P<0.05)。在IR及IP组内,不同时点HIF-1α蛋白表达两两比较差异均有统计学意义(P<0.05);除24 h外其他各时间点IP组大鼠HIF-1α蛋白表达明显高于IR组,差异均有统计学意义(P<0.05)。IR组和IP组大鼠肝组织的HIF-1α蛋白表达与AI、TNF-α、IL-10均呈正相关(r=0.624,0.470,0.312,P<0.05)。结论 IP通过上调HIF-1α蛋白表达,抑制细胞凋亡,减少促炎性因子的释放,增加抗炎性因子的表达,对大鼠肝IR损伤有明显的保护作用。Objective To investigate the protective effects of ischemic preconditioning（ IP） on the injury of hepatic ischemia reperfusion（ IR） in rats and its influence on the expression of hypoxia-inducible factor 1α（ HIF-1α） protein. Methods One hundred and twenty Sprague-Dawley rats were divided into sham operation group（ SO group）,IR group and IP group, with forty rats in each group. In SO group,the hepatoduodenal ligaments of rats were only pulled and separated,and then abdomens were closed. In IR group,the hepatoduodenal ligaments of rats were blocked for 30 minutes and then removed the blocking. In IP group the rats were treated with ischemia for 5 minutes,followed by reperfusion for 5 minutes before the hepatoduodenal ligaments were blocked for 30 minutes. Ten rats of IR group and IP group were sacrificed at 2,6,12 and 24 hours after reperfusion respectively,and the liver tissues were taken out. The liver tissues of rats in SO group were taken out at 2 hours after operation. The levels of aspartate aminotransferase（ ALT）,alanine aminotransferase（ AST）,tumor necrosis factor-α（ TNF-α） and interleukin-10（ IL-10） were measured,and the expression level of HIF-1α protein was detected by immunohistochemical method,the apoptosis of liver cell was detected by TdT-mediated dUTP Nick-End Labeling（ TUNEL） method,and the apoptosis index（ AI） was calculated. Results After reperfusion,the levels of ALT,AST,TNF-α,IL-10 in IR group and IP group at each time point were significantly higher than those in SO group（ P 0. 05）. Compared with IR group,the TNF-α levels were significantly lower and IL-10 levels were significantly higher in IP group at each time point（ P 0. 05）. The levels of ALT and AST at 2,6,12 hours in IP group were significantly lower than those in IR group（ P 0. 05）. The AI in IR group and IP group were significantly higher than those in SO group（ P 0. 05）; the AI in IP group was significantly lower than that in IR group at each time point（ P 0. 05）. The

关 键 词：缺血预处理 肝脏缺血再灌注损伤 缺氧诱导因子1Α 凋亡 

分 类 号：R657.3[医药卫生—外科学]