新生儿缺血缺氧性脑病ceRNA调控网络构建分析  

Construction analysis of ceRNA regulatory network in neonatal hypoxic encephalopathy

作　　者：毕思童 侯礼轩 李丽华[1] BI Sitong;HOU Lixuan;LI Lihua(Beijing Luhe Hospital,Capital Medical University,Beijing 101100)

机构地区：[1]首都医科大学附属北京潞河医院,北京101100

出　　处：《北京生物医学工程》2024年第4期361-369,共9页Beijing Biomedical Engineering

基　　金：首都医科大学2019年度科技技术,社科计划项目(KM201910025001)资助。

摘　　要：目的 分析并构建新生儿缺血缺氧性脑病(hypoxic-ischemic encephalopathy,HIE)竞争性内源RNA(competing endogenous RNA,ceRNA)的调控网络,筛选预测缺血缺氧性脑病诊治的潜在靶点。方法 通过TCGA和GEO数据库获取HIE的非编码RNA高通量测序数据,并进行差异基因分析。根据ceRNA理论构建出circRNA(circular RNA)-miRNA-mRNA网络,进行京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)信号通路和基因本体理论(Gene Ontology,GO)分享,探索和功能注释具有ceRNA潜在功能的circRNA。结果 对GSE121178数据集进行筛选,获得差异circRNAs165个,其中上调59个,下调106个;对GSE181127数据集进行筛选,获得差异miRNAs 63个,其中上调60个,下调3个。用miRecords、miRTarBase、TarBase数据库对获得差异miRNAs进行靶点mRNA预测,并对3个数据库预测结果取交集,共获得11个靶点mRNA(PIK3R1、BACE3、VEGFA、PPP2R2A、LATS2、WDR82、VEZT、TUBA1A、THBS1、SLC7A6、CDK6)。经阈值筛选,筛选了10个共表达circRNAs、4个共表达miRNAs和11个共表达mRNAs,构建circRNA-miRNA-mRNA网络。KEGG富集分析显示:HIE涉及的20个交集蛋白在信号通路中,较为靠前的10条信号通路包括T细胞受体信号通路、Ras信号通路、Rap1信号通路、PI3K-Akt信号通路、MAPK信号通路、FoxO信号通路、ErbB信号通路、胰岛素抵抗、酪氨酸激酶抑制作用、细胞衰老和细胞周期等靶点。说明这些信号通路或与HIE的发生有关。动物实验表明:模型组大鼠神经功能缺损评分高于空白组(P<0.001)。模型组miR-31-5p的mRNA表达水平明显低于空白组(P<0.05),模型组中miR-520g-3p,miR-30a-5p,miR-29a-3p表达水平明显高于空白组(P<0.05);WB显示,模型组中CDK6、THBS1和TUBA1的表达水平明显高于空白组(P<0.05),模型组中VEZT和WDR82的表达水平明显低于空白组(P<0.05)。结论 构建了HIE的circRNA-miRNA-mRNA调控网络,找到了疾病的潜在治疗靶点,为HIE的诊治提供�Objective The regulatory network of competitive endogenous RNA(ceRNA) in neonates with hypoxic-ischemic encephalopathy(HIE) is analyzed and constructed to screen potential targets for diagnosis and treatment of HIE.Methods The high-throughput sequencing data of HIE non-coding RNA were obtained through TCGA and GEO databases,and differential gene analysis was performed.The circRNA-miRNA-mRNA network was constructed according to ceRNA theory.We shared the Kyoto Encyclopedia of Genes and Genomes(KEGG) signaling pathway and Gene Ontology theory(GO),explored and functionally annotated circRNAs with the potential functions of ceRNA.Results The GSE121178 dataset was screened,and 165 circRNAs differences were obtained,of which 59 were up-regulated and 106 were down-regulated.After screening the GSE181127 dataset,63 differential miRNAs were obtained,of which 60 were up-regulated and 3 were down-regulated.Target mRNA was predicted by miRecords,miRTarBase and TarBase databases,and the prediction results of the three databases were intersected to obtain a total of 11 target mRNA(PIK3R1,BACE3,VEGFA,PPP2R2A,LATS2,WDR82,VEZT,TUBA1A,THBS1,SLC7A6,CDK6).After threshold screening,10 co-expressing circRNAs,4 co-expressing miRNAs and 11 co-expressing mRNAs were screened,and the circRNA-miRNA-mRNA network was constructed.KEGG enrichment analysis showed that the 20 intersection proteins involved in HIE were in the signaling pathway.The top 10 signaling pathways included T cell receptor signaling pathway,Ras signaling pathway,Rap1 signaling pathway,PI3K-Akt signaling pathway,MAPK signaling pathway,FoxO signaling pathway,ErbB signaling pathway,insulin resistance,and resistance to tyrosine kinase inhibition,cellular senescence and cell cycle were concentrated.These signals maybe related to the occurrence of HIE.The experimental results showed that the neurological deficit score of the model group was higher than that of the blank group(P<0.001).The level of miR-31-5p in model group was much lower than that in blank group(P<0.05),while the

关 键 词：新生儿 缺血缺氧性脑病 竞争性内源RNA 差异基因分析 

分 类 号：R318[医药卫生—生物医学工程]