家兔脑死亡供肝细胞凋亡水平的定量研究  

The Quantitative Research in Hepatocyte Apoptosis Level from Rabbit Donors after Brain Death

作　　者：钟自彪[1] 张萌[1] 熊艳[1] 王彦峰[1] 叶啟发[1,2] 

机构地区：[1]武汉大学中南医院,武汉大学肝胆疾病研究院,武汉大学移植医学中心,移植医学技术湖北省重点实验室,湖北武汉430071 [2]中南大学湘雅三医院,卫生部移植医学工程技术研究中心,湖南长沙410013

出　　处：《中国普外基础与临床杂志》2016年第1期22-27,共6页Chinese Journal of Bases and Clinics In General Surgery

基　　金：国家自然科学基金委员会-新疆联合基金重点支持项目(项目编号:U1403222);湖北省自然科学基金项目(项目编号:2013CFB258)

摘　　要：目的通过建立稳定的家兔脑死亡模型,应用TUNEL、免疫印迹和免疫组织化学方法检测脑死亡状态下肝脏细胞凋亡水平,从而研究家兔脑死亡供肝细胞凋亡情况。方法 30只新西兰家兔采用缓慢间断颅内加压的方法建立稳定的脑死亡模型,根据脑死亡时间不同分为2、6及8 h组,每组10只家兔,同时设立对应时间点的假手术组(30只)。分别进行相应处理后收集肝脏组织标本,用TUNEL法检测肝细胞凋亡率,用免疫印迹法检测细胞凋亡蛋白Cleaved-caspase 3的表达,用免疫组织化学方法检测Cleaved-caspase 3的表达。结果 TUNEL检测结果显示,脑死亡组各时间点肝细胞凋亡率均高于对应时间点的假手术组(P<0.05),且随着脑死亡时间的延长肝细胞凋亡率逐渐升高(P<0.05)。免疫印迹检测结果及免疫组织化学法检测结果均显示,Cleaved-caspase 3蛋白的相对表达量随着脑死亡时间的延长逐渐增高(P<0.05),且脑死亡组各时间点Cleaved-caspase 3蛋白的相对表达量均高于对应时间点的假手术组(P<0.05)。结论脑死亡供肝与细胞凋亡的关系密切;随着家兔脑死亡时间的延长,肝细胞凋亡水平逐渐增加,从而影响脑死亡供肝的质量。Objective After establishing the rabbit brain death model, TUNEL, western blotting, and immuno- histochemical methods were used to detect hepatocyte apoptosis to study hepatocyte apoptosis level from rabbit donors after brain death. Methods Sixty healthy male New Zealand rabbits were divided into brain death group （n=30） and sham group （n=30）. The rabbits of brain death group were established by increasing intracranial pressure in a modified, slow, and intermittent way, collecting liver tissues after corresponding treatment respectively, using TUNEL to detect apoptosis rate, western blotting and immunohistochemical methods to detect the expression of Cleaved-caspase 3. Results The hepatocyte apoptosis rate at each time point of brain death group were higher than those of the corresponding time point of sham group （P〈0.05）, and the rate of hepatocyte apoptosis increased gradually with the extension of brain death time （P〈0.05）. The results of Western blot assay and immunohistochemistry assay showed that the relative expression amount of Cleaved-caspase 3 protein increased gradually with the extension of brain death time （P〈0.05）, and relative expression amount of Cleaved-caspase 3 protein at each time point of brain death group were higher than those of the corresponding time point of sham group （P〈0.05）. Conclusions The relationship between brain death donor liver and cell apoptosis is closely related. Along with the extension of the brain death time in rabbits, the level of apoptosis of liver cells gradually increased, which affects the quality of liver donors after brain death.

关 键 词：脑死亡 肝脏 细胞凋亡 

分 类 号：R657.3[医药卫生—外科学]