Abstract

Objective

To study the feasibility of plasma extracellular vesicles (EVs) miRNAs as diagnostic biomarkers for Parkinson’s disease (PD).

Methods

Plasma EVs were isolated from 30 PD patients and 30 age- and sex-matched healthy controls. Plasma EVs miRNAs were analysed by qRT-PCR. SH-SY5Y cells were induced by different concentrations of 1-Methyl-4-phenil-pyridinium (MPP⁺) to obtain PD cellular model. The levels of miRNAs and α-synuclein (α-syn) in PD cellular model were analysed by qRT-PCR and Western blot. Receiver operating characteristic (ROC) curve analysis was performed to determine the diagnostic usefulness of the miRNAs in plasma EVs for PD. The gene ontology (GO) and KEGG pathways of the target genes of miRNAs were analysed by softwares.

Results

The level of hsa-miR-30c-2-3p in plasma EVs was significantly higher in PD patients than that in controls, and the levels of hsa-miR-15b-5p, hsa-miR-138-5p, hsa-miR-338-3p, hsa-miR-106b-3p and hsa-miR-431-5p in plasma EVs were lower in PD patients than that in controls. When compared with the control group, the area under the curve (AUC) values for hsa-miR-15b-5p, hsa-miR-30c-2-3p, hsa-miR-138-5p, hsa-miR-431-5p, hsa-miR-338-3p and hsa-miR-106b-3p were all greater than 0.6. The target genes of hsa-miR-15b-5p, hsa-miR-30c-2-3p, hsa-miR-138-5p and hsa-miR-338-3p were enriched in dopaminergic synapse and PD pathway.

Conclusions

The hsa-miR-15b-5p, hsa-miR-30c-2-3p, hsa-miR-138-5p, hsa-miR-106b-3p, hsa-miR-338-3p and hsa-miR-431-5p may be used as potential biomarkers for the diagnosis of PD, and the combined diagnostic accuracy of hsa-miR-15b-5p, hsa-miR-30c-2-3p, hsa-miR-138-5p and hsa-miR-106b-3p was better. The target genes of hsa-miR-15b-5p, hsa-miR-30c-2-3p, hsa-miR-138-5p and hsa-miR-338-3p may regulate the expression of dopamine by dopaminergic synapse and PD pathway.

• Highlights

• Isolation and identification of plasma EVs.

• The miRNAs in plasma EVs may be used as potential biomarkers for the diagnosis of PD.

• When SH-SY5Y cells were induced by different concentrations of MPP⁺, the levels of miRNAs and α-syn changed gradually.

• The target genes of miRNAs were enriched in dopaminergic synapse and PD pathway.

• The target genes of miRNAs may regulate the expression of dopamine.

中文翻译：

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帕金森病患者血浆细胞外囊泡中miRNA的差异表达及意义

摘要

客观的

研究血浆细胞外囊泡 (EVs) miRNAs 作为帕金森病 (PD) 诊断生物标志物的可行性。

方法

从 30 名 PD 患者和 30 名年龄和性别匹配的健康对照中分离出血浆 EV。通过qRT-PCR分析血浆EVs miRNAs。不同浓度的1-甲基-4-苯基吡啶鎓(MPP ⁺ )诱导SH-SY5Y细胞获得PD细胞模型。通过qRT-PCR和Western印迹分析PD细胞模型中miRNA和α-突触核蛋白（α-syn）的水平。进行接受者操作特征 (ROC) 曲线分析以确定血浆 EV 中 miRNA 对 PD 的诊断有用性。通过软件分析miRNA靶基因的基因本体（GO）和KEGG通路。

结果

PD患者血浆EVs中hsa-miR-30c-2-3p的水平显着高于对照组，hsa-miR-15b-5p、hsa-miR-138-5p、hsa-miR- PD 患者血浆 EV 中的 338-3p、hsa-miR-106b-3p 和 hsa-miR-431-5p 低于对照组。与对照组相比，hsa-miR-15b-5p、hsa-miR-30c-2-3p、hsa-miR-138-5p、hsa-miR-431-5p 的曲线下面积 (AUC) 值，hsa-miR-338-3p和hsa-miR-106b-3p均大于0.6。hsa-miR-15b-5p、hsa-miR-30c-2-3p、hsa-miR-138-5p和hsa-miR-338-3p的靶基因在多巴胺能突触和PD通路中富集。

结论

hsa-miR-15b-5p、hsa-miR-30c-2-3p、hsa-miR-138-5p、hsa-miR-106b-3p、hsa-miR-338-3p 和 hsa-miR-431-5p可用作诊断 PD 的潜在生物标志物，以及 hsa-miR-15b-5p、hsa-miR-30c-2-3p、hsa-miR-138-5p 和 hsa-miR-106b-的组合诊断准确性3p更好。hsa-miR-15b-5p、hsa-miR-30c-2-3p、hsa-miR-138-5p和hsa-miR-338-3p的靶基因可能通过多巴胺能突触和PD通路调节多巴胺的表达。

• 强调

• 等离子电动汽车的分离和鉴定。

• 血浆 EV 中的 miRNA 可用作诊断 PD 的潜在生物标志物。

• ^{当不同浓度的MPP +}诱导SH-SY5Y细胞时，miRNAs和α-syn的水平逐渐变化。

• miRNA的靶基因富集于多巴胺能突触和PD通路。

• miRNA的靶基因可能调节多巴胺的表达。