The Study On Changes Of The Whole Transcriptome Expression And The CeRNA Mechanism In The Subacute Stage Of Spinal Cord Injury

[Objective]Spinal cord injury(SCI)is the spine and corresponding spinal cord damage caused by violence,resulting in abnormal sensation,decreased or disappeared muscle strength below the injury level,and even respiratory failure,incontinence,the patient loses labor,may need a wheelchair,ventilator and long-term care.It bring extremely heavy pain and burden to patients and families.In recent years,the global SCI incidence rate has been increasing,but because of the complexity and uncertainty of the pathophysiological mechanism of SCI,the current treatment of spinal cord injury lacks a completely effective method.At present,related researches in the field of epigenetics such as DNA methylation,non-coding RNA(Non-coding RNA,ncRNA)are gradually increasing,The effect of long non-coding RNA(lncRNA),circular RNA(Cicular RNA,circRNA)and microRNA(Micro RNA,miRNA)on the pathophysiology of spinal cord injury needs further study.In this study,the second-generation RNA-seq high-throughput sequencing technology was used to detect the expression changes of the whole transcriptome in the subacute phase of spinal cord injury in mice,and the bioinformatic analysis was used to obtain the biological processes and signaling pathways involved by differentially expressed mRNA and ncRNA in subacute phase after SCI.In addition,after further analysis this study obtained the competitive endogenous RNA(ceRNA)mechanism of between differentially expressed mRNA and ncRNA.[Method]1.In this project,spinal cord tissue of contusion model was isolated,and RNA-seq high-throughput sequencing technology was used to detect the expression changes of the whole transcriptome on 4th day after spinal cord injury in mice.2.Use DAVID database to obtain the biological processes and signaling pathways that may be conducted by differentially expressed mRNA in subacute phase of SCI;use STRING database and Cytoscape software to further construct a protein interaction network analysis of differential mRNA and filter key genes and perform modules analysis.3.Establish lncRNA-miRNA-mRNA network map to clarify the potential ceRNA regulation relationship in the subacute phase of spinal cord injury in mice..[Result]1.A total of 1657 lncRNAs,34 circRNAs,77 miRNAs and 4177 mRNAs were differentially expressed in the SCI mouse model,of which 1026 lncRNAs were up-regulated,631 lncRNAs were down-regulated;26 circRNAs were up-regulated and 8 circRNAs was down-regulated;68 miRNAs were up-regulated,9 miRNAs were down-regulated;2848 mRNAs were up-regulated,and 1329 mRNAs were down-regulated.2.The results of GO analysis suggest that differentially expressed mRNA is involved in cellular processes,biological regulation,cell fraction,cell membrane,catalytic activity,signal transduction activity,molecular transduction activity and molecular function regulator;KEGG results show that differentially expressed mRNA is mainly enriched to signal transduction Signal transduction and catabolism,cell community,cell growth and death,lipid metabolism.3.After constructing protein interaction network,3 core modules and 6 key differential genes were obtained.4.This study explored and constructed the potential ceRNA mechanism of lncRNA-miRNA-Gfap-201/Vegfa-202,which can be used in the next functional verification experiment..[Conclusion]1.This study detected the expression changes of the whole transcriptome in the subacute phase after spinal cord injury.2.Revealed the biological functions and signaling pathways that may be involved by differentially expressed mRNA in the subacute phase after spinal cord injury,and obtained 3 important modules and 6 hub genes in the differentially expressed mRNA.3.This subject further explores the ceRNA relationship of lncRNA-miRNA-Gfap-201/ Vegfa-202,and provides targets for regulating secondary injury of spinal cord injury.