Bioinformatics Analysis Of Differential Gene And LncRNA Expression During Recombinant Human Erythropoietin Treatment Of Spinal Cord Injury In Rats

Background:Spinal cord injury(SCI)is a global health problem,and its incidence is between 15 and 40 million.With the development of economic levels in countries around the world,the incidence of SCI has been increasing year by year.SCI is the most serious complication of spinal injury and it usually causes severe neurological damage and often causes severe dysfunction of the limbs below the spinal cord injury segment.Society places a huge economic burden.Therefore,it is necessary to adopt new treatment strategies to promote the functional recovery of patients with SCI and to better understand the molecular mechanisms of SCI in order to help develop new treatment methods.In previous research,we reported that recombinant human erythropoietin(EPO)reduced apoptosis after SCI in rats and promoted spinal cord function recovery.And as far as we know,there is no research on the molecular mechanism of EPO repairing SCI.Long non-coding RNA(lncRNA)is an RNA transcript longer than 200 nucleotides,which lacks the ability to encode proteins.With the deepening of research,people have discovered that lncRNA is an important regulatory molecule in the human genome,and they play their biological control role in various ways.At the same time,with the development of high-throughput sequencing technology,more and more IncRNAs have been identified,and a recent study also showed that IncRNA imbalance is an important factor in various neurological pathologies and may be involved in SCI play a key role.However,the regulatory mode of IncRNA in EPO-treated damaged spinal cord tissue is currently unclear.Exploring the regulatory mode of LncRNA in SCI and repair processes has some advantages in promoting the advancement of spinal cord treatment methods and the development of specific drugs for spinal cord injury Great significance.Therefore,in this subject,we used high-throughput sequencing to detect differential genes and IncRNA expression profiles in the spinal cord tissue of rats after SCI and EPO treatment,respectively,to establish differential gene expression profiles,and then perform GO analysis and KEGG enrichment analysis on the differential genes and IncRNA.Finally,Cytoscape(vesion 3.2.0)software was used to draw a regulatory network map,and important genes were found.Preliminary analysis of the biological functions regulated by these differential genes and IncRNA was performed.Methods:1.Construction of rat spinal cord injury model:SD rats were randomly divided into three groups:sham operation group,simple SCI group,and EPO-treated SCI group.Rat spinal cord model was constructed according to previous studies.2.Screening of differential gene expression profiles in rats with spinal cord injury:Total RNA from spinal cord tissue was extracted from rat SCI models,and differentially expressed genes and lncRNA profiles of sham operation group,SCI alone group,and EPO-treated spinal cord injury group were detected by chip sequencing At the same time,real-time quantitative PCR experiments were used to verify the expression data of differentially expressed genes and IncRNA on the chip.3.Preliminary analysis of differentially expressed genes and IncRNA in EPO-treated SCI rats:GO analysis and KEGG enrichment analysis of differential genes using R studio,and protein-protein interaction(PPI)analysis using STING.Finally Cytoscape(vesion 3.6.0)software was used to draw the regulatory network of differentially expressed genes and lncRNA.Results:A model of SCI in rats was successfully performed in this experiment,and the model was evaluated successfully using the BBB scoring system.The real-time quantitative PCR experimental data are consistent with the microarray differentially expressed genes and lncRNA expression data,indicating that the microarray data is valid.In the differential gene screening,compared with the sham operation group,4446 genes were differentially expressed in the SCI group(2,471 mRNAs were down-regulated and 1,975 mRNAs were down-regulated).In addition,99 lncRNA changes were found in the injury group(45 lncRNAs were up-regulated,while the other 44 lncRNAs were down-regulated).Compared with the SCI group,the EPO treatment group identified 228 differential genes and found that 14 lncRNAs were changed.Finally,the differential genes were analyzed by GO analysis and KEGG pathway analysis to obtain the corresponding enrichment analysis data,and 10 hub genes were obtained by Cytoscape software analysis.Conclusion:SCI can lead to differential expression of lncRNA and genes.EPO treatment can change the expression profile of differentially expressed lncRNA and genes.These results may provide new insights into the pathophysiology of SCI and the treatment of EPO,and provide a research basis for the development of new treatments.