| Objective: In this study,gene expression profiles and microRNA expression profile data of GENE EXPRESSION OMNIBUS(GEO)in ischemic stroke were screened for differentially expressed genes and differentially expressed microRNAs.Bioinformatics analysis of the function,so as to explore the pathogenesis of ischemic stroke and disease-related genes and microRNAs,molecular mechanisms of its research;provides reference for molecular diagnosis and individualized treatment of ischemic stroke.Methods:1)Gene Spring GX11.5 software was used to screen the differentially expressed genes of gene expression profile data GSE22255.Western-Blot method to verify the expression of differentially expressed genes,and PPI network,GO and KEGG analysis.2)Gene Spring GX11.5 software was used to screen differentially expressed microRNAs of microRNA expression data GSE55937.Real-time PCR was used to validate differentially expressed microRNAs.Targetscan,mi RDB and mi Records were used to predict the target genes.Finally,GO and KEGG analysis was performed.Results:1.Differentially expressed genes Gene expression data GSE22255(1)screened 15 differentially expressed genes.(2)Randomly selected TNF and IL1 B for Western-Blot validation,the results suggest that the same results with the gene chip.By constructing a protein(3)-protein interaction network,IL8(CXCL8),TNF,IL1 B,PGS2,and JUN were shown as network central nodes.(4)The differentially expressed genes were analyzed by gene ontology and we found that these differentially expressed genes were mainly concentrated in the extracellular space,mainly involved in intracellular signal transduction,vascular development,leukocyte migration,transcription and receptor binding.(5)The differentially expressed genes were analyzed by KEGG pathway and we found mainly involved TNF signaling pathway,NF-KB signaling pathway,Toll-like receptor signaling pathway,Nod-like receptor signaling pathway,MAPK signaling pathway.2.Differentially expressed microRNA(1)microRNA expression profile data GSE55937 screened up 6 mirRNA,down 5 mi RNA.(2)Randomly selected mi R-122 and mi R-145 Real-time PCR validation,the results suggest that the same results with the gene chip.(3)The differentially expressed mi RNAs were respectively imported into Target Scan,mi RDB and mi Records databases and intersected to predict 698 target genes,285 up-regulated and 413 down-regulated.(4)Upregulation of target genes were analyzed by KEGG pathway and we found mainly involved TGF-β signaling pathway,Wnt signaling pathway,MAPK signaling pathway.The major biological pathways involved in down-regulation of target genes mainly involve adipocytokine signaling,ECM-receptor interaction,and vascular smooth muscle contraction.Conclusion:1,Microarray date mining methods can be screened for differentially expressed genes and microRNAs associated with ischemic stroke.2,Bioinformatics found that MAPK signaling pathway may be related to the development of ischemic stroke. |
