Isolation, Identification And Analysis Of MicroRNA Expression Profile Of Plasma Exosomes On Patients With Parkinson's Disease

MicroRNAs(miRNAs)are small non-codingRNAs,which have been identified as post-transcriptional regulators by affecting mRNA translation and/or stability.In recent years,growing studies showed that miRNAs participated in the development of Parkinson's disease,and some of the differentially expressed miRNAs were helpful for the diagnosis of PD.Objective:To analysis the expression of plasma exosomal miRNA between the patients with PD and controls,followed by target gene prediction and KEGG pathway analysis,to find some meaningful miRNAs.Methods:1 ? Plasma samples were obtained from 13 PD patients and 13 age/gendermatched healthy individuals.2?Exosomes were isolated from plasma by Invitrogen kit.To ensure isolated exosomes,we used q Nano to analyze their size distribution and particle quantification,and detected their expression of exosomal marker(CD63?Alix)by Western blot.3?We used high-throughput sequencing to profile miRNAs and identified different miRNAs between the two groups.4?MiRNA target gene prediction software were used to predicit the target genes of mi R-195,mi R-485-5p,mi R-15 b and mi R-10a-5p,then KEGG pathway was performed to analysis their target genes.Results:1 ? We obtained particles with mode diameter <100nm and exosomal marker(CD63?Alix),defining as exosomes.2?Compared with controls,the miRNAs(Ratio>2.0,P<0.05)of patients with PD were mainly up-regulated(65.3%).3?The target genes of mi R-10a-5p,mi R-485-5p,mi R-15 b and mi R-195 were all involved in neurotrophin signaling pathway.Conclusions:1?Exosomes were successfully isolated from plasma.2?Plasma exosomal mi R-10a-5p,mi R-485-5p,mi R-15 b and mi R-195 may act as a biomarker for PD.