低氧预适应对大鼠肝癌肝切除术后残肝缺血再灌注中细胞凋亡的影响  

Effect of hypoxic precondition on apoptosis of hepatic ischemia reperfusion with the hepatectomy of transplanted hepatic cancer models in SD rants

作　　者：韩效帆[1] 张培建[1] 李鹏飞[1] 王健[1] 薛同敏[1] 

机构地区：[1]扬州大学第二临床医学院普通外科研究室,扬州225002

出　　处：《国际外科学杂志》2014年第1期18-23,F0003,共7页International Journal of Surgery

基　　金：江苏省扬州市社会发展科技攻关项目基金资助项目（No.YZ2010087）

摘　　要：目的 探讨低氧预适应诱导热休克蛋白70（HSP70）在大鼠肝癌肝切除术后对残肝细胞凋亡的影响及可能的作用机制.方法 采用SD大鼠移植性肝癌模型,根据随机表将60只模型鼠分为A组（假手术组）,B组（缺血状态下肝叶切除组）,C组（低氧预适应＋缺血状态下肝叶切除组）,每组20只.C组的处理方式：术前给予10％氮氧混合气体处理90 min,Pringle法完全阻断入肝血流15 min,同时切除肿瘤所在的肝左外叶.各组分别于术后1,8,12,24 h取残余肝脏标本,用Western blotting免疫印迹法检测HSP70、Bax蛋白的表达,应用流式细胞仪检测肝细胞凋亡率,下腔静脉取血检测ALT、AST,并在光镜下观察各组组织形态学改变,对相关数据进行统计学处理.结果 （1）与A、B组比较,C组检测的各同时相点HSP70的相对量表达明显增加,组间比较差异有统计学意义（均P＜0.05）,A、B组肝组织中仅有微量HSP70表达,两组比较差异无统计学意义（P＞0.05）;（2）与A组比较,B组各同时相点Bax蛋白表达水平明显升高,肝细胞凋亡率明显增加,两组比较差异有统计学意义（均P＜0.01或0.05）,而C组以上各项指标的变化均较B组明显减弱（均P ＜0.05）;（3）C组肝功能较B组改善明显,病理示A组肝组织形态基本正常,C组呈轻微病理学改变,而B组肝组织损伤明显加重.结论 低氧预适应对缺血条件下肝癌肝切除术产生的缺血再灌注损伤具有保护作用,其可能的机制之一是通过上调HSP70蛋白的表达,而HSP70通过降低Bax的表达,抑制细胞凋亡,从而发挥肝脏保护作用.Objective To investigate the effect of Heat shock protein 70 induced by hypoxic precondition on apoptosis of the remnant livers after the hepatectomy of transplanted hepatic cancer models in SD rants and it＇s possible mechanisms.Methods Sixty SD rats of transplanted hepatic cancer models were divided into group A （sham operation group）,group B （IR ＋ hepatectomy group）,and group C （hypoxia precondition ＋ IR ＋ hepatectomy group）,with 20 rats in each group.C group was given 10％ oxygen atmosphere for 90 minutes before operation,In 15 minutes of hepatic vascular occlusion with pringle method,Left lateral of liver which tumor was located was resected.At 1,8,12,24 hours after operation,Five rats of each group were killed respectively,then the remnant liver tissues were sampled to measured the hepatocytes apoptosis rate with flow cytometry,expression of heat shock protein 70 （HSP70） and Bax were detected by Western blotting.The serum AST and ALT activities were measured by an automatic analyzer.The histological changes of the hepatocytes were also observed by optical microscopy.All the data were statistically analyzed.Results （1） Compared with A and B group,the HSP70 expression increased obviously in C group at each time point （all P ＜ 0.05）.The protein showed a weak expression in group A and B group,while the index had no significant differences between two groups （P ＞ 0.05）.（2） In B group compared with A group,the expression of Bax and the hepatocytes apoptosis rate were significantly increased （all P ＜ 0.01 or P ＜0.05）.However,amplitudes of the above changes in C group were significantly lower than those of B group （all P ＜ 0.05）.（3）The liver function was improved in group HP.The pathological examination showed that the liver tissues in A group were general normal and in C group were showed mild changes,but in B group presented evident pathological changes.Conclusions HP can protect against ischemia-reperfusion injury after the hepatectomy of transplanted

关 键 词：低氧预适应 肝肿瘤 再灌注损伤 HSP70热休克蛋白质类 细胞凋亡 

分 类 号：R735.7[医药卫生—肿瘤]