microRNA在大鼠肝移植急性排斥反应中的表达  

microRNA expression in the acute rejection of liver transplantation in rats

作　　者：钟克波[1] 杨定华[1] 李湘竑[1] 林梁[1] 何华[1] 梁建坤[1] 刘潇[1] 梁博[1] 李嘉璐[1] 

机构地区：[1]南方医科大学南方医院肝胆外科,广州510515

出　　处：《中华肝胆外科杂志》2013年第10期771-776,共6页Chinese Journal of Hepatobiliary Surgery

摘　　要：目的探讨microRNA（即miRNA）在大鼠肝移植急性排斥反应中的表达情况。方法以Kamada“双袖套”法行大鼠原位肝移植术（ROLT），分为排斥组（Lewis／BN，n=12）、无排斥组（Lewis／Lewis，n=12）。术后第7d采用miRNA芯片技术确定外周血miRNA表达谱，采用生物信息学方法筛选出表达差异显著的特定miRNA，进而利用qRT-PCR技术对ROLT术后第3、5、7d该特定miRNA进行定量分析，同时结合病理、肝功能结果进行动态化对比分析。结果miR-206在排斥组与无排斥组的表达差异最为显著。排斥组外周血miR-206表达从术后第3d既显著高于无排斥组（t=8．875，P=0．001），且表达量随术后时间延长而逐步上升（F=32．154，P〈0．01）。而两组的排斥反应活动指数（RAI评分）到术后第5d、肝功能指标（TBil、ALT）到术后第7d才出现显著差异。外周血和移植肝中miR-206表达量与RAI评分呈强正相关（rs分别为0．812及0．881，P〈O．01）。在排斥组，外周血与移植肝中miR-206表达量呈正相关（rp=0．644，P=0．024）。结论对于大鼠肝移植急性排斥反应的诊断，外周血特定miRNA检测具有早期、灵敏、特异、稳定等优势，是一种优于病理检查和肝功能化验的诊断方法。【Objective To investigate the role of miRNA expression in the acute rejection of liver transplantation in rats. Methods The rat orthotopic liver transplantation （ROLT） was performed using the two-cuff technique described by Kamada with modification. Rats were randomly divided into two groups, a rejection group （n= 12） and a control group （n= 12）. MicroRNA microarray technology was used to screen the microRNA expression profile in peripheral blood on the 7th postoperative day, and the bioinformatics microRNA screening method detected that miR-206 had the biggest difference in expression level. Peripheral blood samples were gathered before, 3 days, 5 days, and 7 days after ROLT to analyze the expression level of miR-206 quantitatively with qRT-PCR technology. Dynamic contrast analysis was made between the quantitative results of miR-206 and the results of pathological examination and liver function tests. Results There was no significant difference in miR-206 expression level between the rejection group and control group before the operation （t=0. 105, P=0. 921）. MiR-206 expression was significantly higher in the rejection group than in the control group 3 clays after ROLT （t= 8. 875, P = 0. 001）. The expression level of miR-206 in the rejection group also increased gradually over time （F=32. 154, P〈0.01）. However, pathological changes and liver function index did not show obvious differences until the 5th and 7th day postoperatively. The expression of miR-206 in peripheral blood and graft was strongly and positively correlated with the rejection active index （RAI） of acute liver rejection pathology （0. 812 and 0. 881 respectively, P〈0.01）. In the rejection group, the miR-206 level in peripheral blood and graft were positively associated with each other. Conclusions Detection of miR-206 in peripheral blood was significantly better than pathological examination and liver function test in the diagnosis of acute rejection of rat liver transplantation. Moreover, miR-206 was an early s

关 键 词：1microRNA 肝移植 大鼠 急性排斥反应 

分 类 号：R657.3[医药卫生—外科学]