宫内发育迟缓新生小鼠胰腺发育转录组分析

Transcriptome analysis of pancreas development in intrauterine growth retardation neonatal mice

目的:通过全转录组测序比较宫内发育迟缓(intrauterine growth retardation,IUGR)新生鼠与正常新生鼠胰腺长链非编码RNA(long noncoding RNA,LncRNA)及信使RNA(messenger RNA,mRNA)表达谱的差别,探讨IUGR胰岛发育障碍及发生成年糖尿病的潜在机制。方法:建立IUGR新生鼠及正常新生鼠模型,对两种小鼠胰腺进行全转录组测序,分别对LncRNA和m RNA数据进行基因本体论(gene ontology,GO)分析,基于京都基因与基因组百科全书数据库(Kyoto encyclopedia of genes and genomes,KEEG)进行富集分析,全转录组数据联合分析后挑选与胰岛发育相关、表达差异高的LncRNA进行验证。结果:IUGR新生鼠和正常新生鼠差异表达LncRNA 294个,其中上调83个,下调211个;差异表达mRNA 2 000个,其中1 711个上调,289个下调。LncRNA靶基因及差异m RNA的GO显示:生物学途径(biological process,BP)均集中于细胞过程、生物调节及代谢过程;细胞组件(cellular component,CC)集中在细胞及器官等过程;分子功能(molecular function,MF)集中于整合、催化活性、转运活性等。KEEG富集分析显示差异表达的LncRNA靶基因及mRNA主要集中于PI3K-Akt通路、MAPK通路及Foxo通路上。对差异表达LncRNA Snhg12、Rian的验证显示,它们在IUGR鼠与正常鼠中存在明显表达差异,与测序结果一致,且受糖浓度调节。结论:本研究分析并部分验证了IUGR差异表达的LncRNA和m RNA,有助于进一步探讨IUGR新生鼠胰岛发育障碍及发生成年糖尿病的潜在机制及LncRNA在其中的作用。

Objective:This study compared the expression spectrum of long noncoding RNA(lncRNA)and messenger RNA(mRNA)in pancreatic tissue between intrauterine growth retardation(IUGR)and normal neonatal mice,and then explored the potential mechanism of islet developmental disorders in IUGR mice and development of diabetes later in their adulthood. Methods:IUGR and normal neonatal mice models were established,whole transcriptome sequencing in pancreas of two groups were performed,and then gene ontology(GO)and Kyoto Encyclopedia of gene and genomes(KEEG)enrichment analysis were conducted. To further analyse the whole transcriptome sequencing information,we selected several lncRNAs with high differential expression for validation,which were also reported to be involved in islet development. Results:Total 294 lncRNAs were differentially expressed between IUGR and normal neonatal mice,among which 83 were up-regulated and 211 were down-regulated. There were 2 000 differentially expressed mRNAs,of which 1 711 were up-regulated and 289 were down-regulated. The GO analysis of lncRNA target genes and differential mRNAs showed:biological pathways(BP)are concentrated in cellular processes,biological regulation and metabolic processes;cell components(CC)are concentrated in cell and organ processes;molecular function(MF)mainly focuses on integration,catalytic activity,transportation activity,etc. KEEG enrichment analysis highlights the involvement of PI3 K-Akt pathway,MAPK pathway and Foxo pathway. Through verified experiments,the selected lncRNA Snhg12 and Rian showed significant differences between IUGR mice and normal mice,which is consistent with the sequencing results,and the expression were regulated by glucose concentration. Conclusion:This study analyzed and partially verified the lncRNA and mRNA differentially expressed in IUGR,which is helpful to explore the potential mechanism of islet developmental disorders and later adult diabetes in mice born with IUGR,especially the role of lncRNAs.