山羊miR-141靶基因预测及生物信息学分析  

Target Gene Prediction and Bioinformatics Analysis of miR-141 in Capra hircus

作　　者：马建青 宋鹏琰 杨清芳 孔建军 宋占锋 张亚丽 吴东蔚 徐增年 赵宁 周荣艳[6] 吴占勇 MA Jianqing;SONG Pengyan;YANG Qingfang;KONG Jianjun;SONG Zhanfeng;ZHANG Yali;WU Dongwei;XU Zengnian;ZHAO Ning;ZHOU Rongyan;WU Zhanyong(Department of Orthopedic Laboratory,North China Medical&Health Group Xingtai General Hospital,Xingtai 054000,China;College of Animal Science and Technology,Tarim University,Alaer 843300,China;Xingtai Agriculture and Rural Affairs Bureau,Xingtai 054001,China;Third Hospital of Hebei Medical University,Shijiazhuang 050051,China;Deparment of Zoology,Hebei Medical University,Shijiazhuang 050017,China;College of Animal Science and Technology,Hebei Agricultural University,Baoding 071001,China)

机构地区：[1]华北医疗健康集团邢台总医院骨科研究所,邢台054000 [2]塔里木大学动物科学与技术学院,阿拉尔843300 [3]邢台市农业农村局,邢台054001 [4]河北医科大学第三医院,石家庄050051 [5]河北医科大学实验动物学部,石家庄050017 [6]河北农业大学动物科技学院,保定071001

出　　处：《中国畜牧兽医》2024年第10期4211-4221,共11页China Animal Husbandry & Veterinary Medicine

基　　金：邢台市重点研发计划项目(2023ZC156);河北省自然科学基金(H2021108006)。

摘　　要：【目的】探究miR-141在动物机体糖脂代谢和骨代谢中的调控机制,为糖尿病合并骨质疏松症动物模型构建及相关基因表达调控机制研究提供依据。【方法】利用miRbase数据库获取不同物种miR-141成熟序列,通过BioEdit及WebLogo软件进行序列比对及保守性分析;利用PROMO及JASPAR在线软件对山羊miR-141启动子区转录因子结合位点进行预测;利用TargetScan和miRwalk数据平台对miR-141靶基因进行预测;利用DAVID和KOBAS在线软件对靶基因进行GO功能和KEGG通路富集分析;利用Networkanalyst数据库建立miRNA靶基因蛋白互作网络并绘制网络调控图;通过YM500V2在线数据库分析miR-141在山羊不同组织中的表达情况。【结果】miR-141成熟序列在不同物种间高度保守;山羊miR-141启动子区分布有转录因子C/EBPα、MyoD、YY1和Sp1等结合位点;选用不同数据库预测到Nfatc2、Egr2和Fasl等89个靶基因。GO功能富集分析发现,靶基因主要富集在RNA聚合酶Ⅱ启动子转录调控与转录DNA模板等生物过程、细胞核与细胞质等细胞组分、蛋白结合与DNA结合等分子功能。KEGG通路富集显示,靶基因主要富集在肌动蛋白细胞骨架的调节、细胞紧密连接、轴突引导、Hedgehog信号通路、cGMP-PKG信号通路与癌症中的microRNA等。miR-141靶基因蛋白互作分析显示,Nfatc2和Egr2基因由Fasl基因连接,构成与其他蛋白相对复杂的靶向关系,这些关键基因参与了与机体糖脂代谢及骨代谢相关的TGF-β、Hedgehog及Wnt/β-catenin等多条信号通路。miR-141在山羊多种组织中均有表达,其表达水平具有组织差异性。【结论】miR-141可通过靶向Nfatc2、Egr2和Fasl基因参与调节Hedgehog及Wnt/β-catenin等多条信号通路,并进一步调控山羊糖脂代谢、肌肉骨骼系统生理生化反应。【Objective】This study was aimed to explore the regulatory mechanism of miR-141 in glucolipid metabolism and bone metabolism,and provide a basis for establishing animal model related to diabetes mellitus with osteoporosis and researching the regulation mechanism of miR-141 related gene expression.【Method】Mature miR-141 sequences of different species were obtained from miRbase database for sequence alignment and conservative analysis used BioEdit and WebLogo softwares.The PROMO and JASPAR online softwares were used to predict the transcription factor binding site of miR-141 in Capra hircus.The target genes of intersection were predicted by TargetScan and miRwalk online tools.GO function and KEGG pathway enrichment analysis of the predicted target genes were performed by DAVID and KOBAS online softwares.The functional protein interaction was analyzed with Networkanalyst database,meanwhile drawing the network regulation diagram.The YM500V2 online database was used to analyze the expression of miR-141 target genes in different tissues of Capra hircus.【Result】miR-141 mature sequences among different species were highly conservative.There were many transcription factor binding sites such as C/EBPα,MyoD,YY1 and Sp1 in the promoter region of miR-141 in Capra hircus.A total number of 89 target genes included Nfatc 2,Egr 2 and Fasl were predicted using different databases.GO function enrichment analysis results showed that the predicted target genes of miR-141 mainly enriched in the regulation of transcription from RNA polymeraseⅡpromoter and regulation of transcription-DNA-templated for biological process,nucleus and cytoplasm for cellular component,protein binding and DNA binding for molecular function.KEGG pathway enrichment analysis results showed that the target genes were significantly enriched in regulation of actin cytoskeleton,tight junction,axon guidance,Hedgehog signaling pathway,cGMP-PKG signaling pathway,microRNA in cancer and other signaling pathways.miR-141 target genes interaction analysis res

关 键 词：山羊 miR-141 靶基因 糖脂代谢 骨代谢 

分 类 号：S827[农业科学—畜牧学] Q78[农业科学—畜牧兽医]