癫痫大鼠海马组织的转录组学特征分析

An analysis of transcriptomic characteristics of hippocampal tissue in epileptic rats

目的探究癫痫的转录组学特征,寻找癫痫诊治的潜在靶点。方法选取雄性幼龄SD大鼠17只,采用抽签法随机将大鼠分为2组:对照组(8只)、CSRS组(9只)。基于氯化锂-匹罗卡品构建的慢性自发性癫痫(CSRS)大鼠模型,取CSRS大鼠及对照组大鼠海马组织进行转录组测序(RNA-seq),并对信使RNA(mRNA)及微小RNA(miRNA)进行差异分析、信号通路富集分析及差异表达蛋白互作用网络构建寻找核心调控基因。结果对照组及CSRS组的mRNA进行差异分析获得的354个差异表达基因(DEGs),信号通路富集分析DEGs与离子转运调控、组织发育、应激反应等相关。蛋白互作网络分析发现WDR88、SHANK2、TEC、NFKBIZ、EPHA8等为DEGs中主要的核心调控基因。同时获得差异表达miRNA,并进行靶基因预测及GO通路富集分析,发现靶基因主要与离子型谷氨酸受体复合物、谷氨酸受体活性、谷氨酸盐化的钙离子通道活性、NF-κB诱导激酶活性等分子功能相关。结论基于RNA-seq发现差异基因表达异常是癫痫发生的重要原因,是导致神经元发生异常的、同步化放电的关键因素。

Objective To investigate the transcriptomic characteristics of epilepsy and explore potential targets for the diagnosis and treatment of epilepsy.Methods A total of 17 young male Sprague-Dawley rats were randomly divided intocontrol group(n=8)and chronic spontaneous recurrent seizures(CSRS)group(n=9).Lithium chloride-pilocarpine was used to establish a rat CSRS model.Transcriptome sequencing(RNA-seq)was performed on the rat hippocampal tissues from the CSRS and control groups.A differential analysis and a signaling pathway enrichment analysis on messenger RNA(mRNA)and microRNA(miRNA)were performed,and a differentially expressed protein-protein interaction network was constructed to search for core regulatory genes.Results A total of 354 differentially expressed genes(DEGs)were obtained via the differential analysis on mRNA between the control group and the CSRS group.The signaling pathway enrichment analysis showed that DEGs were related to ion transport regulation,tissue development,stress response,etc.The protein-protein in teraction network analysis revealed that WDR88,SHANK2,TEC,NFKBIZ,EPHA8,etc.,were the main core regulatory genes in DEGs.In addition,differentially expressed miRNA was obtained.Target genes were predicted,and the GO path-way was enriched.It was found that the target genes were mainly related to the molecular functions such as ionic glutamate receptor complex,glutamate receptor activity,glutamate-dependent calcium channel activity,and NF-κB-inducing kinase activity.Conclusions Based on the results of RNA-seq,it was found that abnormal expression of differentially expressed genes is an important cause of epilepsy,and it is a key factor leading to abnormal and synchronized discharge of neurons.