肠组织长链非编码RNA表达谱在新生儿坏死性小肠结肠炎组织中的变化

Changes of long non-coding RNA expression profiles in intestinal tissues from neonatal necrotizing enterocolitis

目的:长链非编码RNA(lncRNA)参与新生儿坏死性小肠结肠炎(necrotizing enterocolitis,NEC)的发生和发展,本研究探讨新生儿NEC肠组织lncRNA表达谱的变化及其潜在机制。方法:收集需要手术治疗的NEC患者肠样本11份,同期非NEC需要手术新生儿肠样本7份。采用Illumina HiSeq对6份肠样本(对照组3份,NEC组3份)进行lncRNA高通量测序,得到NEC组和对照组的lncRNA差异表达谱。剩余部分标本采用实时聚合酶链反应检测差异表达的lncRNA的相对表达量,验证高通量测序的结果。对差异表达的基因进行Gene Ontology(GO)分析和KEGG信号通路分析。结果:对照组和NEC组组间存在显著差异的lncRNA有5257个。表达上调的NONHSAG008675.3,NONHSAG020715.2,NONHSAG038187.2以及下调的NONHSAG028744.3的验证结果与测序结果相符。NEC肠组织DUOX2,IL-6,TNF,SAA1表达上调。对表达水平有显著差异的lncRNA靶基因进行GO富集,GO富集的生物进程主要有对刺激的调节、分子黏合和功能的调节、信号网络及转录信号调节等方面。KEGG主要富集PI3K-Akt,炎症性肠病,NF-κB等。结论:LncRNAs可能参与NEC的发病机制,炎症相关的lncRNAs可能是关键影响因素之一。

Objectives:Long non-coding RNAs(lnc RNAs)were involved in the development and regulation of necrotizing enterocolitis(NEC)in premature infants.To investigate the changes of lnc RNA expression profile in intestinal tissues of NEC for its possible mechanisms.Methods:Intestinal samples were collected from 11 patients with NEC who needed surgery(the NEC group),and 7 from neonatal non-NEC patients with surgery(the Control group).Lnc RNA’s changes in intestinal samples(3 in the Control group and 3 in the NEC group)were analyzed with high-throughput sequencing.Part of the remaining samples were detected by real-time polymerase chain reaction(real-time PCR),and the results were used to validate the results of high-throughput sequencing.Gene Ontology(GO)analysis and KEGG signaling pathway analysis were performed on differentially expressed genes.Results:There were 5257 different lnc RNAs between the control group and the NEC group.The results of up-regulated lnc RNAs(NONHSAG008675.3,NONHSAG020715.2,NONHSAG038187.2)and down-regulated lncR NA(NONHSAG028744.3)were confirmed to be consistent with the results of high-throughput sequencing.Expressions of DUOX2,IL-6,TNF,and SAA1 were up-regulated in intestinal tissues of NEC.GO analysis showed that the different lnc RNAs were involved in regulation of stimulation,molecular junction and function,and signal transduction and transcription.KEGG analysis identified mainly biological pathways involved in inflammatory bowel disease,PI3K-Akt,NF-κB,etc.Conclusion:Lnc RNAs might be involved in the pathogenesis of NEC and the inflammation-related lncRNAs may be one of the key factors.