| BackgroundAtrial fibrillation(AF)is one of the most common clinical arrhythmia at present in China,the prevalence of AF increases with age.The total prevalence of AF in China is about 0.77%,and prevalence rates can be as high as 7.5%in people over the age of 80 years.AF not only affects the quality of life of patients,but also causes complications such as thromboembolism and heart failure in severe cases,one of the most serious and harmful complications is stroke.However,the mechanism of AF occurrence and maintenance is still unclear.Long non-coding RNA(lncRNA)is a kind of non-coding RNA with a length of more than 200 nucleotides.An increasing number of studies have shown that lncRNA is involved in the occurrence and development of various cardiovascular diseases,including AFObjectiveWe established a goat atrial fibrillation model and used second-generation sequencing to explore the biological significance of lncRNA-mRNA network in the occurrence and maintenance mechanism of AF.MethodsFourteen male adult goats weighing between 16 and 24 kg were randomly divided into two groups,AF group(n=7)and Normal group(n=7),We made a goat atrial fibrillation model by continuous rapid pacing of the left atrium and compared the atrial effective refractory(AERP)period before and after pacing end point in each group.Total RNA was extracted from the left atrial appendage tissue of the goats.Agilent Bioanalyzer 2100 was used for total RNA quality testing,NanoDrop One spectrophotometer,and Qubit(?)3.0 Fluorometer for quantitative and purity testing of total RNA.The expression profiles of lncRNAs and mRNAs in each group were detected by RNA-seq,high-throughput sequencing technology,and lncRNAs and mRNAs with different expressions were screened out after pretreatment of the original data.The differentially expressed lncRNAs and mRNAs were verified by qPCR.Weighted correlation network analysis(WGCNA),Gene Ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG)and other methods were used to analyze the expression profile to explore the possible correlation mechanism in the occurrence and maintenance of AFResults1.Goat atrial fibrillation model was successfully constructed,and AERP in AF group was significantly shorter after pacing end point than before pacing(p<0.05).2.LncRNA and mRNA were differentially expressed in AF group compared with Normal group.Among them,21 lncRNA were differentially expressed,15 were up-regulated and 6 were down-regulated(cutoff power:p<0.05,|Fold Change|>1.5).However,differences in mRNA expression were 1079,757 were up-regulated and 322 were down-regulated(cutoff power:p<0.05,|Fold Change|>1.5).3.QPCR verified ENSCHIT00000009973(lncRNA),ENSCHIT00000004773(lncRNA)and ENSCHIG00000011256(mRNA),and the results were consistent with the expression of RNA-seq,all of which were up-regulated.4.WGCNA analysis identified 23 gene modules,and modules Blue,Darkred,Royalblue were significantly related with AF.All the genes in the Blue module were analyzed by GO analysis and KEGG analysis,and they were mainly enriched in oxidation-reduction process,oxidoreductase activity,MAPK signaling pathway,etc.Selected the Blue module analysis with the closest correlation,and found 22 core genes.Further,Cytoscape software tool was used to observe that MSTRG.4358.1 in the visual network was highly correlated with 5 core genes,namely ATP5F1B,GHITM,IDH3B,SLC25A4 and MUL1.ConclusionMSTRG.4358.1 may be involved in the occurrence and maintenance of AF by regulating the five hub genes ATP5F1B,GHITM,IDH3B,SLC25A4 and MUL1,laying the foundation for related functional verification in the next step. |
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