| Objective:Ischemic stroke(IS)is a common cerebrovascular disease and the main cause of death and disability in the world,especially in developing countries.The biomarkers related to IS are mainly various proteins related to pathological processes.However,gene expression variations are currently mainly used for early diagnosis and prediction of diseases,due to the instability of traditional biomarkers.Long non-coding RNA(lnc RNA),as an epigenetic modifier,is widely involved in gene transcription regulation.It regulates the occurrence and development of diseases through co-expression with its targeted m RNA.This study intends to provide a theory basis for the study of biological functions and regulatory mechanisms in the pathophysiological process of IS by identifying the expression profiles of lnc RNA and m RNA in IS,as well as the gene expression regulation methods they participate in disease and the body's metabolic regulation pathways that affect disease occurrence.Methods:Three patients with first diagnosed ischemic stroke(A1,A2,A3)and 3 controls(D1,D2,D3)in the Second Hospital of Jilin University from July 2017 to December 2017 were selected.Collect information such as age,smoking,drinking,and related chronic disease prevalence,and retain 3ml of fasting venous blood the next morning after hospitalization.Using TRIzol reagent to extract RNA for high-throughput sequencing.Using Ballgown software to analyze the differential expression of all lnc RNA and m RNA,using the Pearson correlation coefficient method to analyze the correlation between lnc RNA and m RNA,and performing gene ontology(GO)enrichment analysis of differential lnc RNA target genes through the R software GO seq package.To enrich KEGG(Kyoto Encyclopedia of Genes and Genomes)via KOBAS,and build protein-protein interaction network based on STRING database.Using Cytoscape software to draw network visualization graphics,and use Clue GO to construct GO/KEGG pathway functional classification network for differentially expressed m RNA.Results:1.High-throughput sequencing results show that there are 5 differentially expressed lnc RNAs(4 up-regulated and 1 down-regulated)and 144 differentially expressed m RNAs(70 up-regulated and 74 down-regulated)related to ischemic stroke;2.The Lnc RNA-m RNA co-expression network contains 946 nodes and 1009 lines.Lnc RNA ENST00000574212.1 has the most co-expressed m RNA,and is the most important lnc RNA in the network,constituting the core regulation of the co-expression network Module3.Enrichment analysis of GO entries of target genes that differentially express lnc RNA yielded 166 GO entries that were significantly enriched,including 113 Biological process entries,34 Cellular component entries,and 19 Molecular function entries;4.Herpes simplex infection,Influenza A,RIG-I-like receptor signaling pathway and m TOR signaling pathway are significantly enriched in KEGG pathway enrichment analysis of differentially expressed lnc RNA target genes;5.The protein interaction network that differentially expresses lnc RNA target genes contains a total of 417 nodes and 810 edges.EP300 and NFKB1 are the two most important factors that regulate protein expression;6.The three genes HLA-DQB1,HLA-DQA1 and HLA-DRB5 in the human leukocyte antigen(HLA)family are the key genes that overlap in the Clue GO pathway network that differentially expresses m RNA.Conclusions:1.Five differentially expressed lnc RNAs related to ischemic stroke may participate in the regulation of the mechanism of ischemic stroke by affecting the body's immune and inflammation pathways;2.EP300 and NFKB1 are core regulatory factors that significantly related to ischemic stroke,and regulate the interaction of protein expression,which is the key content of future molecular biological mechanism research;3.HLA family genes related to ischemic stroke may mainly mediate Th17 cell differentiation pathway,inflammatory bowel disease pathway and asthma pathway to participate in the regulation of IS pathogenesis. |
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