摘要
旨在通过粪肠球菌感染小鼠构建体内血脑屏障损伤模型,从miRNA角度探究小鼠血脑屏障通透性的分子调控机制。试验采用粪肠球菌XJ1感染小鼠构建血脑屏障损伤模型,并通过检测血脑屏障的通透性、脑组织病理切片观察,血清中PCT和CRP的变化判断模型构建结果。将感染组和对照组脑组织样品进行转录组测序,构建miRNA文库,筛选差异表达miRNA,进行qPCR验证。试验成功构建粪肠球菌感染小鼠体内血脑屏障损伤模型,共鉴定22对差异表达的miRNAs,其中上调表达的miRNAs有8个,下调表达的miRNAs有14个。GO分析结果,包括了20个细胞成分、20个生物学过程和20个分子功能。KEGG富集分析表明,差异表达miRNA的靶基因显著富集于调节干细胞多能性的信号通路、RAS信号通路、MAPK信号通路、神经胶质瘤、FcεRI信号通路等。实时荧光定量PCR结果表明,差异miRNA表达趋势与测序结果一致。得出差异表达的mmu-miR-34b-5p和mmu-miR-21a-5p是关于血脑屏障通透性改变的候选miRNA,为探究miRNA对血脑屏障的调节提供了理论参考。
An in vivo blood-brain barrier damage model was constructed to infect mice with Enterococcus faecalis to explore the molecular regulation mechanism of blood-brain barrier permeability in mice from the perspective of miRNA.Enterococcus faecalis XJ1 was used to infect mice to construct a blood-brain barrier damage model,and the results of the model construction were judged by detecting the permeability of the blood-brain barrier,observation of pathological sections of brain tissue,and changes in serum PCT and CRP.The brain tissue samples of the infection group and the control group were subjected to transcriptome sequencing,a miRNA library was constructed,and differentially expressed miRNAs were screened and verified by qPCR.A blood-brain barrier damage model in mice infected with Enterococcus faecalis was successfully constructed.A total of 22 pairs of differentially expressed miRNAs were identified,of which 8 miRNAs were up-regulated and 14 were down-regulated.GO analysis results included 20 cellular components,20 biological processes and 20 molecular functions.According to the KEGG enrichment analysis,the target genes of differentially expressed miRNAs were significantly gathering in the regulation of stem cell pluripotency pathways such as signaling pathway,RAS signaling pathway,MAPK signaling pathway,glioma,FcεRI signaling pathway,etc.The results of real-time fluorescence quantitative PCR showed that the differential miRNA expression trends were consistent with the sequencing results.The differentially expressed mmu-miR-34b-5p and mmu-miR-21a-5p are candidate miRNAs for the permeability changes of the blood-brain barrier,providing a reliable theoretical reference for exploring the regulation of miRNA on the blood-brain barrier.
作者
陈明杰
李益涛
曹梦园
王晨豫
闫雪琪
陈杰
齐亚银
CHEN Mingjie;LI Yitao;CAO Mengyuan;WANG Chenyu;YAN Xueqi;CHEN Jie;QI Yayin(School of Animal Science and Technology,Shihezi University,Shihezi,Xinjiang 832000,China)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2022年第11期2202-2210,共9页
Chinese Journal of Veterinary Science
基金
国家自然科学基金资助项目(31260610,31760737)。
关键词
粪肠球菌
miRNA
血脑屏障
高通量测序
Enterococcus faecalis
miRNA
blood brain barrier
high throughput sequencing
