Sequencing Study Of CircRNA In Peripheral Blood From Parkinson’s Disease

Objective: Sequencing technology was analyzed to the differential expression of circRNA in peripheral blood of patients with Parkinson’s disease and normal controls.The differential expression levels of circRNA in patients with Parkinson’s disease and normal controls were compared to explore the significant difference circRNA and its possible signal pathways and corresponding target genes.The study provides a new vision for the discovery of diagnostic markers of Parkinson’s disease by depth understanding of the role of circRNA in the pathogenesis of Parkinson’s disease.Methods: The study subjects were grouped to Patients with Parkinson’s disease（n = 4）and normal controls（n = 4）.Peripheral venous blood was collected from two groups,and RNA was extracted and purified according to the PAXgene Blood RNA Kit operating manual.RNA concentration and integrity were detected by NanoDrop 2000 And Agilent 2100.Sequencing technology was used to analyze the expression of mRNA and circRNA.Heat map and volcano map were used to select the mRNA and circRNA.Signal pathways and target genes related to Parkinson’s disease development are obtained through GO analysis and KEGG analysis.The miRTarBase database software was used to predict the binding sites of circRNA and miRNA.The TargetScan database software was used to predict the binding sites of mRNA and miRNA.In order to study the biological function of circRNA in Parkinson’s disease,the ceRNA network was constructed by predicting the binding sites of mRNA,miRNA and circRNA.Besides,qRT-PCR was further verified to the significantly differently expressed circRNA.Results: Sequencing analysis was performed by comparing the RNA expression profiles of patients with Parkinson’s disease and healthy controls.The present study indicated that 766 mRNAs and 411 circRNAs were expressed differentially between the PD and NC.The results show that 766 differentially expressed mRNAs（273mRNAs were upregulated,and 493 mRNAs were downregulated）and 411 differentially expressed circRNAs（129 circRNAs were upregulated,and 282 circRNAs were downregulated）.Through KEGG enrichment analysis of mRNA,it was found that the expression pathway of Parkinson’s disease was the second.Differentially expressed mRNA in patients with Parkinson’s disease was associated with mitochondrial respiratory chain dysfunction.The competing for endogenous RNA（ceRNA）network includes 10 differentially expressed mRNAs,13 predicted miRNAs,and 10 differentially expressed circRNAs.The expression of certain mRNAs and circRNAs was found to be associated with mitochondrial defects and abnormal respiratory chains in differentially expressed mRNAs and circRNAs.Eight significant differentially expressed circRNAs were verified using qRT-PCR,of which hsa_circ₀009190 was statistically different（P<0.05）.Conclusion: There are a large number of differentially expressed circRNAs in peripheral blood of patients with Parkinson’s disease.The sponge effect of circRNA combined with miRNA may play an important role in the regulation of Parkinson’s disease gene expression.According to GO,KEGG enrichment and ceRNA network analysis,it was found that mitochondrial respiratory chain dysfunction was closely related to Parkinson’s disease.The qRT-PCR verification indicated that hsa_circ₀009190 may be related to the occurrence and development of Parkinson’s disease.