Profile Of MiRNA In Hippocampus Of Epileptic Mice And Its Bioinformatics Analysis

Background Epilepsy is a common disease of the nervous system.There are about 50 million epilepsy patients worldwide.As a chronic disease whose etiology is not yet clear,its long-term and repeated seizures have caused huge physical and psychological problems to patients and their families.,economic burden.Micro RNAs(miRNAs)are a class of endogenous non-coding small RNAs with regulatory functions,which degrade target m RNAs by inducing the formation of silencing complexes,or repress the translation of target m RNAs.Recently,more and more literatures have reported that miRNAs play an extremely important role in the signal transduction,differentiation and maturationcentra,and is closely related to various diseases.Objective To explore the changes of hippocampal miRNA expression profile by establishing an animal model of lithium chloride pilocarpine status epilepticus,and to carry out bioinformatics analysis on the miRNAs differentially expressed compared with normal mice,so as to find possible related signal pathways or epilepsy mechanisms.Methods(1)Twenty-six clean grade healthy adult C57BL/6 mice(weighing 20-30g)were selected and divided equally into epilepsy group and control group.The epilepsy group mice were injected intraperitoneally with lithium chloride pilocarpine to establish a status epilepticus(SE)model,and animal behavior was observed and recorded.(2)Extract miRNA from the hippocampus of two groups of mice,analyze the miRNA expression profile of the two samples using RNA-Seq technology,and conduct statistical comparison to find differences.(3)Process and filter sequencing results,and conduct bioinformatics analysis of the results.(4)Target gene prediction software was used to predict the target genes of the target miRNA,and gene clustering(GO)and KEGG databases were used to analyze the biological significance and biological pathways of the target genes,respectively.Result(1)Differently expressed miRNAs: In mice with status epilepticus,a total of 55 miRNAs were differentially expressed compared to the normal group,with 38 levels up and17 levels down.(2)The most significant differences were identified among the miRNA5(mmu-let-7a-1-3p、mmu-let-7a-2-3p、mmu-let-7c-5p、mmu-let-7d-5p、mmu-let-7e-5p).(3)Target gene prediction: Input the information of 5 miRNAs and use the target gene prediction software Targetscan to match the known target gene m RNA connection sites to obtain the target genes targeted by these differentially expressed miRNAs.(4)Predict possible new miRNAs(novel＿189,novel＿190,novel＿194,novel＿221,novel＿229)through miREvo and mirdeep2 software.(5)Bioinformatics analysis: Input the obtained target genes into the Metascape software for bioinformatics analysis to obtain the Gene Ontology project and the KEGG biological signal pathway.Looking at the annotated information and past research data on these target genes and biological pathways,we found that they point to mechanisms such as neural synaptic remodeling,endogenous apoptosis,and actin remodeling.Conclusion(1)The differentially expressed miRNAs in status epilepticus mice were measured by high-throughput sequencing technology.Among them,55 miRNAs were statistically significant,of which 38 were up-regulated and 17 were down regulated.(2)Five miRNAs with the most significant differences were screened using statistical tools.Through the bioinformatics tools such as Targetscan,GO,and KEGG,the target genes corresponding to five miRNAs were predicted,and it was found that these miRNAs might be related to GTPase signal transduction,GTPase activity regulation,cell morphology regulation,endometrial tissue and transcriptional regulation,and also related to MAPK signaling pathway,actin pathway,and cell cycle pathway.