大鼠腹部手术应激后血清miRNA表达谱的差异分析

目的:应用miRNA芯片技术建立大鼠腹部手术应激后血清miRNA差异表达谱，从中发现与早期手术创伤后应激相关的miRNAs。方法:建立大鼠部分肝切除手术模型。检测手术前后大鼠血清中ALT、AST和CRP浓度水平及肝脏病理改变，评估腹部创伤后大鼠应激情况。采用miRNA表达谱芯片筛选手术前后大鼠血清中差异表达的miRNAs，并采用实时荧光定量RT-PCR(real-time RT-PCR)进行验证。采用生物信息学软件(MiRand和TargetScan)预测候选miRNA的靶基因。结果:血清miRNA表达谱出现明显改变，24个miRNAs在2/3部分肝切除术（PH）后24h大鼠血清中表达明显升高，特别是miR-9，其在2/3PH术后24h大鼠血清中的表达水平为术前的50多倍，real-time RT-PCR检测miR-9的结果与芯片结果相符。通过生物信息学预测，CDH1、E-cadherin、MTHFD2、PDYN、MCPIP1、BCL2L11、CMA1、Map3k1等可能是miR-9的靶基因。结论:腹部手术创伤后，大鼠血清表达谱发生明显变化，提示miRNA可能参与调控创伤后应激反应。

Altered expression levels of miRNAs in rat serum post abdominal operation

Objective:To screen and identify the differential expression of miRNAs in rat serum post traumatic injury by the miRNA microarray technique.Methods:We chose partial hepatectomy (PH) as traumatic injury model.The serum ALT，AST，CRP and histological changes of the liver were measured to evaluate the severity of trauma.The miRNA microarray was used to assess the level and composition of serum miRNAs.The target genes of candidate miRNA were predicted by bioinformatics software (MiRand and TargetScan).Results:There was significantly differential expression of miRNAs in rat serum post traumatic injury (24 miRNAs were more than 2-fold up-regulated).Especially，the expression of miR-9 showed the highest up-regulated (more than 50-fold).The results showed that CDH1，E-cadherin，MTHFD2，PDYN，MCPIP1，BCL2L11,CMA1，Map3k1 may be the target genes of miR-9.Conclusion:Injury caused by abdominal operation could make significant change in the spectra and levels of serum miRNAs，suggesting that miRNAs may be involved in regulating the stress response post trauma.