Department of Endocrinology，the first Affiliated Hospital of Nanjing Medical University，Nanjing 210009
摘要：目的：对正常孕鼠和宫内发育迟缓(intrauterine growth retardation，IUGR)小鼠孕期胰岛进行高通量测序分析，筛选出差异表达的长非编码RNA（long noncoding RNA，lncRNA）和信使RNA（messenger RNA，mRNA），为深入探讨IUGR孕鼠胰岛功能障碍的发病机制提供理论基础。方法：提取IUGR成年后孕鼠和正常孕鼠胰岛并进行高通量测序，筛选出差异表达的lncRNA和mRNA，进行lncRNA和mRNA关联分析，对lncRNA相关的靶mRNA进行基因本体论(gene ontology，GO)功能注释分析、京都基因和基因组百科全书(kyoto encyclopedia of genes and genomes，KEGG)通路富集分析，重点关注高差异表达且可能影响孕期胰岛功能的lncRNA。结果：IUGR孕鼠(IUGR pregnancy，IP)和正常孕鼠(Normal pregnancy，NP)差异表达的lncRNA1007个，其中483个上调，524个下调；差异表达的mRNA50个，其中22个上调，28个下调。对差异性lncRNA的靶mRNA和差异性mRNA进行功能分析，GO分析示它们参与的生物学途径集中于细胞及组织过程、生物调节、代谢及应激过程；分子功能集中在整合、催化活性、转运活性、分子功能调节等方面；KEGG分析示它们主要集中在代谢、癌症、次生代谢产物的生物合成、PI3K-AKT及MAPK通路。对lncRNA FTX和Neat1进一步分析表明，IP、NP的FTX表达量分别较未孕时下降（P＜0.001），而 Neat1上升（P＜0.01）；FTX、Neat1表达量在IP和NP间有显著差异（P＜0.05），且表达量受糖浓度调节。lncRNA FTX与其靶mRNA均为反式（trans）作用，lncRNA Neat1与Frmd8为顺式（cis）作用，余为trans作用。结论：本研究筛选出IUGR孕鼠和正常孕鼠胰岛中差异表达的lncRNA和mRNA，lncRNA通过不同互作关系调控靶mRNA，调节糖代谢过程，为进一步探究IUGR孕鼠胰岛功能障碍的发病机制提供了新思路。
Abstract：Objective：High-throughput sequencing analysis was performed in adult islets of normal mice and intrauterine growth retardation (IUGR) mice during pregnancy, which screened out differentially expressed long noncoding RNA (lncRNA) and messenger RNA (mRNA) to provide a theoretical basis for further exploring the pathogenesis of islet dysfunction in pregnant mice born with IUGR. Methods: Islets were extracted from IUGR and normal pregnant mice for high-throughput sequencing analysis. The differentially expressed lncRNAs and mRNAs were screened out?and association analysis between them was performed. We conducted gene ontology (GO) analyisis, Kyoto Encyclopedia of gene and genome (KEGG) enrichment analysis on taget mRNAs of differentially expressed lncRNAs. Emphasis was placed on the highly differentially expressed lncRNAs, especially those that may be involved in the regulation of islet function during pregnancy. Results: There were 1007 differentially expressed lncRNAs between IUGR pregnancy (IP) and normal pregnancy (NP) , among which 483 were up-regulated and 524 were down-regulated. 50 mRNAs were differentially expressed, of which 22 were up-regulated and 28 were down-regulated. The GO analysis of differentially expressed lncRNAs’target mRNAs and differentially expressed mRNAs showed that biological process (BP) mainly focused on cellular and tissue processes, biological regulation, metabolism and stress processes; Molecular functions (MF) were concentrated in integration, catalytic activity, transport activity, molecular function regulation,etc. KEGG enrichment analysis highlighted the involvement of metabolic pathway, cancer pathway, biosynthesis of secondary metabolites, PI3K-AKT pathway and MAPK pathway. We furether analysed lncRNA FTX and Neat1. The expression level of FTX in normal mice and IUGR mice during pregnancy decreased (P＜0.001), while LncRNA Neat1 increased (P＜0.01). FTX and Neat1 exhibited significantly difference in the expression between IP and NP (P＜0.05), and both were regulated by glucose concentration. The association between lncRNA FTX and its target mRNAs was trans action. The association between lncRNA Neat1 and Frmd8 was cis action, while the rest were trans action. Conclusions: In this study, differentially expressed lncRNAs and mRNAs in adult islets of normal mice and intrauterine growth retardation (IUGR) mice during pregnancy were screened. Besides, lncRNAs can regulate target mRNAs through different interaction modes, thus influencing glucose metabolism, which provides new ideas for further exploring the pathogenesis of islet dysfunction in pregnant mice born with IUGR.