山西医科大学学报2024，Vol.55Issue(10)：1314-1323,10.DOI:10.13753/j.issn.1007-6611.2024.10.008
基于转录组数据探究MEF2D在癫痫中的作用机制
Role and mechanism of MEF2D in epilepsy based on transcriptomic data
摘要
Abstract
Objective To investigate the role of myocyte enhancer factor 2D(MEF2D)in the pathogenesis of epilepsy(EP).Methods Mouse hippocampal neuronal cell line HT22 was used as the research model.Three distinct gene loci(686,912,and 1283)within MEF2D were targeted for knockdown using lentiviral vectors.The study was divided into four groups:sh686 group,sh912 group,sh1283 group,and shNC group.RT-qPCR and Western blot were performed to verify the successful knockdown of MEF2D.Cells with successful MEF2D knockdown were subjected to apoptosis detection.RNA-seq was performed on the selected cells to identify differen-tially expressed genes(DEGs),regulated alternative splicing events(RASE),and regulated alternative splicing genes(RASGs).Gene ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were per-formed on both DEGs and RASGs.Results RT-qPCR and Western blot results confirmed successful MEF2D knockdown in sh912 group.Flow cytometry analysis revealed a significantly higher rate of early apoptosis in sh912 group than in shNC group(P<0.05).RNA sequencing analysis identified 638 differentially expressed genes(DEGs).GO analysis of DEGs primarily enriched biological pro-cesses related to glycolysis,carbohydrate metabolism,and regulation of redox reactions.KEGG pathway analysis highlighted signifi-cant enrichment in pathways such as Nod-like receptor signaling,tumor necrosis factor(TNF)signaling,hypoxia-inducible factor(HIF)-1 signaling,and IL-17 signaling.A total of 1 265 differentially RASEs were identified in the AS analysis,and intron retention(IR)was the most prevalent type,followed by alternative 5′ splice site(A5SS)and alternative 3′ splice site(A3SS).GO analysis of RASGs predominantly enriched biological processes related to DNA repair,RNA splicing,and protein transport.KEGG pathway analysis of RASGs highlighted significant enrichment in pathways such as Rap1 signaling,PI3K/Akt signaling,and phosphatidylinosi-tol signaling.A total of 28 genes were identified as overlapping between DEGs and RASGs.Conclusion Knockdown of MEF2D could promote the apoptosis of hippocampal neurons.Intron retention is the main alternative splicing event that occurs after MEF2D knockdown.MEF2D may participate in the apoptosis response by affecting cellular functions such as glycolysis and redox reactions.关键词
癫痫/MEF2D/差异表达基因/选择性剪切/内含子保留/细胞凋亡/RNA测序Key words
epilepsy/MEF2D/differentially expressed gene/alternative splicing/IntronR/apoptosis/RNA sequence分类
医药卫生引用本文复制引用
韩登峰,焦腾飞,热娜·阿不都萨拉木..基于转录组数据探究MEF2D在癫痫中的作用机制[J].山西医科大学学报,2024,55(10):1314-1323,10.基金项目
新疆维吾尔自治区自然科学基金项目(2022D01C759) （2022D01C759）
