基于mRNA高通量测序筛选调控小鼠术后谵妄重要差异基因

Screening of Important Differential Genes Regulating Postoperative Delirium in Mice Based on mRNA High-Throughput Sequencing

目的基于高通量测序技术筛选小鼠海马组织中术后谵妄(POD)发生的相关重要差异基因。方法将16只健康雌性C57小鼠随机分为术后谵妄(POD组)和对照组(CON组),每组8只。采用异氟烷麻醉下剖腹探查构建POD模型,术后多时间点行为学检测POD改变。于造模后24 h收集小鼠海马组织进行高通量测序,对显著差异基因进行基因本体(GO)分析及京都基因和基因组百科全书(KEGG)通路数据库分析,采用RT-qPCR验证关键差异表达基因(DEGs)。结果相比于对照组,POD组共鉴定了105个DEGs,其中79个DEGs上调,26个DEGs下调;GO分析结果显示,主要富集的生物过程包括应激反应、代谢过程等,主要表达于细胞器、细胞膜,参与的分子过程涉及转录活性调节等。KEGG分析提示,通路主要富集于NF-κB、趋化因子、mTOR、FoxO和Hippo等信号通路;综合上述结果并结合RT-qPCR验证筛选出Cdkn1a、Nfkbia、Card14、Wnt3、Bcl2、Gng11、Grm3和Akt28个关键基因。结论高通量测序筛选出的差异表达基因,可能通过NF-κB、趋化因子、mTOR等信号通路介导POD发生。

Objective To screening of important differential genes related to the occurrence of postoperative delirium(POD)in mouse hippocampus based on high-throughput sequencing technology.Methods 16 healthy female C57 mice were randomly divided into postoperative delirium group(POD group)and control group(CON group),with 8 mice in each group.POD model was constructed by laparotomy under isoflurane anesthesia,and POD changes were detected at multiple time points postoperatively.Mouse hippocampus tissue was collected for high-throughput sequencing 24 hours after modeling,gene ontology(GO)analysis and kyoto encyclopedia of genes and genomes(KEGG) pathway database analysis for significantly different genes,validation of key differentially expressed genes(DEGs)by RT-qPCR.Results Compared with control group,105 DEGs were identified in POD group,of which 79 DEGs were up-regulated and 26 DEGs were down-regulated.GO analysis showed that the main enriched biological processes included stress responses,metabolic processes,which were mainly expressed in organelles,cell membrane,and molecular processes involved in the regulation of transcriptional activity.KEGG analysis showed that pathways were mainly enriched in signaling pathways such as NF-κB,chemokines,mTOR,FoxO and Hippo.8 key genes,such as Cdkn1a,Nfkbia,Card14,Wnt3,Bcl2,Gng11,Grm3 and Akt2,were screened out by combining the above results and RT-qPCR validation.Conclusion Deferentially expressed genes screened by high-throughput sequencing may mediate the occurrence of POD through signaling pathways such as NF-κB,chemokines,and mTOR.