摘要
目的本研究分别将野生型(WT)、Gper1敲低(Gper1-KD)大鼠海马组织进行转录组测序,探讨G蛋白偶联雌激素受体1(G-protein coupled estrogen receptor 1,GPER1)影响癫痫发病可能的信号通路和分子机制。方法海马组织进行RNA提取和cDNA文库构建,与NCBI数据库大鼠基因组及基因注释比对,通过FPKM值,筛选组别间差异表达基因(DEGs)。将DEGs进行GO富集、KEGG富集分析,构建蛋白互作网络,利用RT-qPCR法对关键DEGs进行验证。结果Gper1-KD组与WT组相比,筛选(Fold change>2且FDR<0.01)后,检测到DEGs 2253个,其中上调基因1380个,下调基因873个;GO结果显示,DEGs主要分布在淋巴细胞趋化性、细胞分泌、巨噬细胞趋化性、中性粒细胞趋化性、血管生成正向调控等生物过程;KEGG结果显示,DEGs相关分子信号通路主要有癌症信号通路、PI3K-Akt通路、癌症蛋白聚糖相关信号通路、细胞黏附相关通路、MAPK信号通路等;RT-qPCR结果表明,Mapk12、Pdpk1、Foxo3、Camk2d、Pik3cg等基因表达水平差异具有显著性。结论MAPK、TNF信号通路在GPER1影响癫痫发生中可能起关键作用,GABA能神经元和Pik3cg在GPER1影响癫痫易感性方面可能发挥重要作用。
Aim To focus on the RNA-Seq analysis of WT,Gper1-KD hippocampal tissues of rats and to seek the possible signal molecule and pathways in epileptogenesis mediated by G-protein coupled estrogen receptor 1(GPER1).Methods RNA was extracted and cDNA library was constructed,then the reliable data were screened and blasted with rat genome from NCBI database.Then the DEGs were selected according to FPKM value.GO enrichment,KEGG enrichment and protein interaction network analysis were performed.Results On the grounds of screening conditions(FC≥1.5,P<0.05)for DEGs,there were 2253 genes selected between WT and Gper1-KD group animals,and 1380 genes were significantly up-regulated and 873 down-regulated.The GO enrichment results revealed DEGs mainly played a part in the biological process including lymphocyte chemotaxis,secretion by cell,macrophage chemotaxis,neutrophil chemotaxis and positive regulation of angiogenesis.Moreover,the KEGG enrichment results demonstrated that the DEGs were mainly involved in pathways in cancer,PI3K-Akt pathway,proteoglycans in cancer,focal adhesion and MAPK pathways.RT-qPCR results showed that the expression levels of Gper1,Foxo3,Camk2d,Pdpk1,Pik3cg,Mapk12 were significantly changed.Conclusions MAPK,TNF signaling pathway may play a key role in GPER1 affecting epileptogenesis,GABAergic neurons and Pik3cg may play an important role in GPER1 influencing epilepsy susceptibility.
作者
左娣
郝世杰
杨盼
李苗
任晓璠
丁娜
马文倩
王盼盼
王诗雨
戎伟芳
刘昆梅
ZUO Di;HAO Shi-jie;YANG Pan;LI Miao;REN Xiao-fan;DING Na;MA Wen-qian;WANG Pan-pan;WANG Shi-yu;RONG Wei-fang;LIU Kun-mei(Ningxia Key Laboratory of Cerebrocranial Diseases,Ningxia Medical University,Yinchuan 750004,China;College of Clinical Medicine,Ningxia Medical University,Yinchuan 750004,China;School of Stomatology,Ningxia Medical University,Yinchuan 750004,China;College of Basic Medical Sciences,Shanghai Jiao Tong University,Shanghai 200025,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2024年第4期709-715,共7页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 82360269)
宁夏重点研发项目(引才专项,No 2021BEB04017)
宁夏自然科学基金资助项目(No 2022AAC03189)
大学生创新创业训练计划(No S202310752055)。
