线粒体途径在缺血后处理减轻大鼠肝脏缺血再灌注损伤作用中的机制研究  

Ischemic postconditioning alleviating liver ischemia-reperfusion injury in rats via mitochondrial pathway

作　　者：林杰[1] 何飞[1] 吴林熹[1] 黄汉飞[1] 曾仲[1] 

机构地区：[1]昆明医科大学第一附属医院,650032

出　　处：《中华器官移植杂志》2012年第10期624-628,共5页Chinese Journal of Organ Transplantation

基　　金：云南省自然科学基金（2007C137M）;云南省科技厅联合基金（200703009R）;云南省社会发展科技计划基金（2008CA026）

摘　　要：目的研究缺血后处理（IPO）减轻大鼠肝脏缺血再灌注损伤（IRI）的机制中关于线粒体途径发挥的作用。方法按随机数字表法将SD大鼠分为3组，IRI组和IPO组建立肝移植模型，IRI组受鼠在供肝植入后，立即开放门静脉恢复血液供应；IPO组受鼠在门静脉完全开放前行IPO，即对门静脉行开放60s、夹闭60S，共重复6次。假手术组大鼠在开腹后仅游离肝周韧带，未进行肝移植。术后6h（IRI组和IPO组为血流完全开放后6h，下同），取各组受鼠的血液和肝组织样本，检测肝功能，采用逆转录聚合酶链反应法检测肝组织B淋巴细胞因子-2（Bcl-2）mRNA和Bcl-2共存蛋白质（Bax）mRNA的表达，采用蛋白质印迹法检测肝组织细胞色素C（Cyt-c）蛋白的表达，使用流式细胞仪检测肝组织的细胞凋亡与坏死率及线粒体跨膜电位的变化，电镜下观察肝细胞线粒体的超微结构。结果与假手术组相比，IRI组和IPO组肝功能明显受损（P〈0．05），肝组织BaxmRNA的表达明显上调（P〈O．05），Bcl-2mRNA的表达明显下降（P〈0．05），Cyt-c蛋白含量显著升高（P〈0．05），肝组织细胞凋亡坏死率明显增加（P〈0．05），肝细胞线粒体跨膜电位明显下降（P〈0．05）。而IP0组以上各项指标的改变均较IRI组明显减轻（P〈0．05），肝细胞线粒体的形态结构改变也明显改善。结论IPO可能通过抑制线粒体途径来减轻大鼠肝移植中肝脏的IRI。Objective To investigate the effect of ischemie postconditioning （IPO） on ischemia/ reperfusion injury via mitochondrial pathway. Methods Male Sprague-Dawley rats were divided into three groups by means of random number table. Rats in IRI group and IPO group received liver transplantation. The portal vein in IRI group was opened immediately after liver implantation to restore the blood supply. The graft rats in IPO group received IPO before the portal vein was completely opened： 60-s ischemia and 60-s reperfusion of the portal vein, repeated 6 times）. The rats in the sham-operation group were only subject to dissociation of the liver ligament. Six h after portal vein reperfusion, liver function was tested. The expression of Bcl-2 mRNA and Bax mRNA was detected by using real time-PCR. The protein expression of Cyt-c was detected by using Western blotting. The apoptosis and necrosis of liver tissue and the mitochondrial transmembrane potential were detected by using flow cytomertry. The changes of mitochondrial structure were observed under an electron microscope. Results The liver functions in IRI group and IPO group were significantly worse （P〈0. 05）, the levels of Bax mRNA were significantly higher （P〈0. 05）, the levels of Bcl-2 mRNA were significantly lower （P〈0. 05）, the levels of Cyt-c protein were significantly higher （P〈 0. 05）, the levels of apoptosis and necrosis were significantly higher （P〈0. 05）, and mitochondrial transmembrane potential （MTP） of liver cells was significantly lower （P〈0. 05） than in sham operation group. The changes of all the parameters in IRI group were more significant than in IPO group （P〈0. 05 ）. The morphological changes of the liver mitochondria were also significantly aggravated in IRI group as compared with IPO group. Conclusion IPO reduced IRI by inhibiting the mitochondrial signaling pathway in rats undergoing liver transplantation.

关 键 词：肝移植 再灌注损伤 缺血后处理 线粒体 

分 类 号：R657.3[医药卫生—外科学]