肝毛细线虫感染小鼠不同时间肝脏差异表达miRNA的鉴定及功能分析

Identification and bioinformatics analysis of deregulated miRNA in liver from mice at different days post-infection with Capillaria hepatica

目的:分析小鼠感染肝毛细线虫后第20和90天肝脏miRNA表达谱,筛选并鉴定差异表达miRNA。方法:将20只6周龄BABL/c雄性小鼠随机分为4组,分别为第20天感染组和对照组、第90天感染组和对照组,感染组以20卵/只的剂量灌胃感染期肝毛细线虫卵。分别于感染后第20和90天取对照组和感染组小鼠肝脏进行miRNA高通量测序,筛选感染后不同时间肝脏差异表达的miRNA,预测其靶基因并进行GO富集分析和KEGG信号通路分析,采用qRT-PCR对7个差异表达的miRNA进行检测。结果:第20天感染组筛选出差异表达的miRNA共81条,第90天感染组差异表达miRNA共122条;第20天和第90天感染组共同差异表达的miRNA有27条,其中均上调21条,均下调6条。差异miRNA靶基因主要参与了信号传导、转录调控等,可能参与的信号通路有Wnt信号通路、Ras信号通路等。qRT-PCR检测结果显示感染组miR-142a-3p、miR-342-3p、miR-34a-5p、miR-150-5p、miR-214-3p表达上调,miR-455-3p和miR-1948-5p表达下调(P<0.05)。结论:小鼠感染肝毛细线虫后肝脏miRNA呈现差异表达,差异表达的miRNA可能参与肝纤维化的进程。

Aim:To analyze the expression profiles of miRNA in the liver of mice infected with Capillaria hepatica on the 20th and 90th day,so as to screen and identify differentially expressed miRNA.Methods:Twenty 6-week-old BABL/c male mice were randomly allocated into 4 groups:the 20th day infection group and the control group,as well as the 90th day infection group and the control group.The infection group was infected with 20 Capillaria hepatica embryonated eggs by gavage.On the 20th and 90th day post-infection,the livers of the mice were harvested and RNA sequencing was used to screen the aberrant miRNAs.The target genes of aberrant miRNAs were estimated.Finally,GO enrichment analysis and KEGG pathway analysis of the target genes were performed.Seven differentially expressed miRNAs were selected for qRT-PCR validation.Results:A total of 81 and 122 differentially expressed miRNAs were identified from the 20th day infection group and 90th day infection group.There were 27 differentially expressed miRNAs in common between the 20th day infection group and the 90th day infection group,among which,21 were up-regulated and 6 were down-regulated.GO analysis of the target genes were mainly involved in the regulation of intercellular signal transduction,DNA transcription and so on.KEGG pathway analysis showed that the enriched pathways were Wnt signaling pathway,Ras signaling pathway,ect.The results of qRT-PCR showed that miR-142a-3p,miR-342-3p,miR-34a-5p,miR-150-5p and miR-214-3p were up-regulated in infection groups,while miR-455-3p and miR-1948-5p were down-regulated(P<0.05).Conclusion:Differential expressions of miRNAs in liver have been observed in mice infected with Capillaria hepatica,and these differentially expressed miRNAs may be involved in the progression of liver fibrosis.