摘要
目的 探索缺氧缺血性脑损伤(hypoxia-ischemic brain damage,HIBD)后新生大鼠肾脏中长链非编码RNA(long non-coding RNA,lncRNA)表达谱的改变.方法 选择健康3日龄新生SD大鼠26只,随机分为HIBD组和正常对照组,HIBD组构建新生大鼠HIBD模型,两组均提取肾脏总RN A,使用HiSeq 2500平台进行高通量测序,用R软件对两组lncRNA表达量进行差异分析.采用实时定量聚合酶链反应(quantitative real-time-PCR,qRT-PCR)验证上调最显著的3个lncRNA和下调最显著的3个lncRNA的表达水平,进一步对差异表达的lncRNA进行靶基因预测,并进行基因本体分析和京都基因与基因组百科全书通路分析,探索靶基因可能的生物学功能.结果 在HIBD组大鼠肾脏样本中共鉴定出2 588个lncRNA,长度主要分布在300～500 bp.差异分析鉴定出差异表达的lncRNA 19个,其中上调13个,下调6个.分别对上调最显著的3个lncRNA(MSTRG.11816、NONRATG011285.2、NONRATG0 13597.2)及下调最显著的 3 个 lncRNA(NONRATG011218.2、NONRATG009165.2、MSTRG.66)进行qRT-PCR验证,结果显示,HIBD组lncRNA NONRATG0 11285.2的相对表达量高于正常对照组,lncRNA MSTRG.66的相对表达量低于正常对照组,差异有统计学意义(P＜0.05).靶基因预测显示,lncRNA NONRATG011285.2的靶基因主要为EIF4E2和FMR1,lncRNA MSTRG.66的靶基因主要为MYC和HRAS.结论 HIBD不仅会影响脑,还会造成肾脏lncRNA改变.
Abstract
Objective To study the changes of long non-coding RNAs(lncRNAs)in the kidneys of neonatal rats after hypoxic-ischemic brain damage(HIBD).Methods Twenty-six healthy 3 day-old Sprague-Dawley(SD)rats were randomly assigned into HIBD group and control group.HIBD model was established according to protocol in HIBD group.Total RNAs were extracted from kidneys in both groups.HiSeq 2500 platform was used for high-throughput sequencing analysis and R software was used to compare the differences of lncRNA expressions.Real-time quantitative polymerase chain reaction(qRT-PCR)was used to verify the top three up-regulated and down-regulated lncRNAs and to predict possible target genes.The biological functions of the target genes were explored using gene ontology(GO)analysis and Kyoto encyclopedia of genes and genomes(KEGG)pathway analysis.Results A total of 2 588 lncRNAs were identified,mostly 300-500 bp long.Differential analysis identified 19 lncRNAs with altered expressions:13 up-regulated and 6 down-regulated.qRT-PCR confirmed the top three up-regulated(MSTRG.11816,NONRATG011285.2,NONRATG013597.2)and down-regulated(NONRATG011218.2,NONRATG009165.2,MSTRG.66)lncRNAs.Comparing with the control group,the HIBD group showed significantly higher NONRATG011285.2 expression and lower MSTRG.66 expression(both P＜0.05).Target gene prediction showed that the target genes of lncRNA NONRATG011285.2 were mainly EIF4E2 and FMR1,the target genes of lncRNA MSTRG.66 were MYC and HRAS.Conclusions HIBD affects both the brain and the kidney,altering kidney lncRNA.
维普
万方数据