大鼠肝脏缺血再灌注损伤对COX-2和micorRNA-101的表达

Impact of Hepatic Ischemia-reperfusion Injury on COX-2 and MicroRNA-101 Expression in Rats

目的探讨大鼠肝脏缺血再关注损伤对COX-2和micorRNA-101表达的影响。方法通过建立大鼠肝脏缺血再灌注损伤模型,并行肝叶切除术,术后通过Quantitative real-time PCR (q RT-PCR)及Western Blot(WB)技术分析COX-2基因在肝脏缺血再灌注过程中的表达情况;通过生物信息学预测可能与COX-2基因相互作用的重要micro RNA,并分析这些micro RNA在肝脏缺血再灌注过程中的表达模式及其对COX-2的表达调控作用。结果在肝脏缺血再灌注后,COX-2基因m RNA及蛋白表达水平显著升高;micro RNA-101在肝脏缺血再灌注过程中表达下调,其表达趋势和COX-2基因表达相反;生物信息预测发现,micro RNA-101可能结合于COX-2m RNA 3'-UTR区域;采用micro RAN-101 mimics处理大鼠永生化肝细胞系BRL-3A细胞可导致COX-2基因表达下调。结论肝脏缺血再灌注可引起micro RNA-101表达下调,进而导致COX-2基因表达上调,并最终影响肝细胞周期改变。

Objective To investigate impact of hepatic ischemia-reperfusion injury on COX-2 and microRNA-101 expression of rats. Methods We established a rat model of hepatic ischemia-reperfusion and performed the Partial hepatectomy. We analyzed the expression of cox-2 by Quantitative real-time PCR (qRT-PCR) and Western Blot (WB);Bioinformatics was used to predict the important microRNAs that may interact with COX-2,and to analyze the expression patterns of these microRNAs during hepatic ischemia-reperfusion and their regulatory effects on COX-2 expression. Results The expression of cox-2 increased significantly after hapatic ischemia reperfusion;the expression of microrna-101 was down-regulated, the mechanism contrary to cox-2;Bioinformatics predicted that microRNA-101 might bind to the 3'-UTR region of COX-2 mRNA,using microran-101 mimics treat the rat liver derived cell line (BRL-3A) can lead to down-regulation of cox-2 gene expression. Conclusions Hepatic ischemia reperfusion can induce the down-regulation of microrna-101 expression, which leads to the up-regulation of cox-2 gene expression, and ultimately affects the changes of hepatic cell cycle.