Differential Expression And Mechanism Analysis Of Long Noncoding RNA After Intracerebral Hemorrhage

Objective: Intracerebral hemorrhage is a severe disease with high mortality and high morbidity.There are very few effective clinical therapies for patients with cerebral hemorrhage.Scientists focus on the mechanism of secondary brain injury after intracerebral hemorrhage and the mechanism of nerve regeneration and repair for a long time.Previous studies have shown that some non-coding RNAs can participate in biological processes such as injury,repair and cell regeneration by regulating gene transcription,translation and post-translational modification.In the development and progression of disease,non-coding RNAs have a significant differential expression.Previous studies focused on the microRNAs in the development of intracerebral hemorrhage more,and research about long chain non-coding very little.This study will analysis the differential expression of lncRNA after cerebral hemorrhage via animal experiments and predict its biological function and mechanism of action analysis.Method: Adult male C57BL/6 mice were randomly divided into intracerebral hemorrhage group(CH group)and sham operation group(C group).According to the stereotactic technique,15 ul autogenous femoral arterial blood was injected into the right basal ganglia of brain to establish the model of intracerebral hemorrhage.In the sham operation group,the other steps were with the group of cerebral hemorrhage in addition to the non injection of autologous femoral artery blood.Mice with successful modeling use the Focal Deficits scale score.Twenty-four hours after modeling,tissue(100mg)around the injection site were taken.100 mg tissue were taken in the sham-operation group at the same place.RNAs were extracted.LncRNA and mRNA in the brain tissue were screened using Arraystar mouse LncRNA V3.0 chip.The differentially expressed lncRNA and mRNA were picked using the Agilent GeneSpring GX v12.1 software.Through GO and KEGG database for biology and pathway analysis.Some of the lncRNAs were selected for qRT-PCR to verify the relative content of lncRNA.TargetScan,miRanda and other databases were used to analyze the mechanism of lncRNA-miRNA-mRNA(ceRNA mechanism),and to predict the biological role of lncRNA-miRNA-mRNA in the pathogenesis of intracerebral hemorrhage.Data statistical analysis using SPSS Statistics 21,GraphPad Prism 7 and other software,measurementdata use t test,enumeration data use χ2 test,P <0.05 was statistically significant.Result: 1.Cerebral hemorrhage model were successful made,hemiplegia appeared in the left side.2.Chips analysis showed that 125 lncRNAs were significantly differentially expressed in brain tissue of mice with intracerebral hemorrhage(Fold Change≥2,P<0.05),74 were up-regulated and 51 were down-regulated.There were 247 mRNA significantly differentially expressed(Fold Change≥2,P <0.05),222 up-regulated and 25down-regulated.3.The results of GO analysis suggested that the three biological processes most relevant to the changes of these m RNAs were as follows: immune response,immune system process and defense response.The most relevant three cell sites were extracellular region,extracellular space,extracellular region part;the most relevant three molecular functions are: endopeptidase inhibitor activity,protein binding,endopeptidase regulator activity.4.Pathway analysis indicated that differentially expressed lncRNAs and mRNAs exerted biological functions through phagosome and intercellular adhesion molecule-related pathways.5.One of the lncRNAs(uc011xxu.1)was verified,the relative expression is 2.17(P <0.05),and it is up-regulated.6.LncRNA-miRNA-mRNA network established by 84 miRNAs,20 mRNA and uc011 xxu.1.7.Among the 20 m RNAs,the most relevant m RNA is Usb 1.It may regulate the endonuclease activity in the nucleus and participate the repairment of cell,RNA metabolism and nitrogen-containing compound metabolism.Conclusion: LncRNAs have significant differential expression in the brain tissue of mice after cerebral hemorrhage.The mechanisms of these lncRNAs in the impair brain tissue are complicated and may become a new therapeutic target and biomarker for patients with cerebral hemorrhage in clinical.