miR-150-5p在非酒精性脂肪性肝纤维化中的表达及靶基因的生物信息学分析

Expression profiling and bioinformatic analysis of miR-150-5p and its target genes in nutritional fibrotic steatohepatitis

目的通过对miR-150-5p进行靶基因预测及生物信息学分析,为miR-150-5p靶基因相关实验验证提供数据支持,并为深入研究miR-150-5p在非酒精性脂肪性肝纤维化中的调控机制及生物学功能提供理论指导。方法将健康雄性C57BL/6J小鼠随机分为2组,模型组10只采用胆碱-蛋氨酸缺乏饲料喂养制备非酒精性脂肪性肝纤维化模型,正常组10只采用胆碱-蛋氨酸充足的饲料喂养,均饲养8周。采用microRNA(miRNA)芯片检测2组小鼠肝组织中miR-150-5p表达水平,选择TargetScan、PicTar及miRanda靶基因预测软件预测miR-150-5p靶基因的交集作为分析的基因集合,分别进行功能富集分析,应用DAVID数据库进行靶基因信号转导通路富集分析。结果模型组小鼠肝组织中miR-150-5p表达水平显著高于正常组(P<0.05),miR-150-5p靶基因集合分别富集在ATP结合、嘌呤核苷酸结合以及蛋白酶的活性等分子功能(P均<0.05),主要参与磷脂代谢、蛋白质氨基酸的磷酸化和去磷酸化、血管再生与重建及促凋亡等生物学过程(P均<0.05),靶基因信号转导通路显著富集于胰岛素信号通路、MAPK信号通路、TGF-β信号通路等信号转导通路中。结论miR-150-5p参与了非酒精性脂肪性肝纤维化发生发展过程,为非酒精性脂肪性肝纤维化新型诊断标志物与治疗靶点的研究提供了理论依据。

Objective It is to provide data support for the experimental verification of miR-150-5p target gene through the target gene prediction and bioinformatics analysis of miR-150-5p,and provide theoretical guidance for the in-depth study on the regulation mechanism and biological function of miR-150-5p in non-alcoholic fatty liver fibrosis.Methods The healthy male C57BL/6J mice were randomly divided into 2 groups.10 mice in the model group were fed with choline-methionine deficient feed to establish the models of non-alcoholic fatty liver fibrosis,and 10 mice in the normal group were fed with choline-methionine sufficient feed,all fed for 8 weeks.The expression level of miR-150-5p in the liver tissues of the mice in the two groups was detected by microRNA(miRNA)chips,and TargetScan,PicTar and miRanda target gene prediction software were selected to predict the intersection of miR-150-5p target genes as the gene set for functional enrichment analysis,DAVID database was used for enrichment analysis of target gene signal transduction pathways.Results The expression level of miR-150-5p in the liver tissue of the model group was significantly higher than that of the normal group(P<0.05).The miR-150-5p target gene set was enriched in molecular function such as ATP binding,purine nucleotide binding and protease activity(all P<0.05),mainly involved in phospholipid metabolism,phosphorylation and dephosphorylation of protein and amino acids,angiogenesis and remodeling,and pro-apoptosis biological processes(P<0.05),significant enrichment of target gene signal transduction pathways in the insulin signal pathway,MAPK signal pathway,TGF-βsignal pathway and other signal transduction pathways.Conclusion MiR-150-5p is involved in the occurrence and development of non-alcoholic fatty liver fibrosis,which providing a theoretical basis for the study of new diagnostic markers and therapeutic targets for non-alcoholic fatty liver fibrosis.