外泌体miR-1246对帕金森病的诊断价值  

Diagnostic value of exosomal miR-1246 for Parkinson’s disease

作　　者：王晓蓓 杨珊[1] 李亚昙[1] 杨新玲[2] WANG Xiaobei;YANG Shan;LI Yatan;YANG Xinling(State Key Laboratory of Pathogenesis,Prevention and Treatment of High Incidence Diseases in Central Asia,Department of Neurology,the Second Affiliated Hospital of Xinjiang Medical University,Urumqi 830063,China;Xinjiang Key Laboratory of Nervous System Disease Research,Xinjiang Medical University,Urumqi 830017,China)

机构地区：[1]省部共建中亚高发病成因与防治国家重点实验室,新疆医科大学第二附属医院神经内科,新疆乌鲁木齐830063 [2]新疆医科大学新疆神经系统疾病研究重点实验室,新疆乌鲁木齐830017

出　　处：《新医学》2024年第11期861-869,共9页Journal of New Medicine

基　　金：省部共建中亚高发病成因与防治国家重点实验室开放课题项目(SKL-HIDCA-2022-NKX3);天山英才科技创新领军人才项目(2022TSYCLJ0066)。

摘　　要：目的探讨外泌体miR-1246对帕金森病(PD)的诊断价值。方法收集2022年9月至2023年4月在新疆医科大学第二附属医院住院的30例PD患者(PD组)和接受体检的30名健康对照者(对照组)的血清样本,运用实时定量PCR分析外泌体中miR-1246的表达水平,使用受试者操作特征(ROC)曲线评估miR-1246在PD诊断中的预测能力。用1-甲基-4-苯基吡啶离子(MPP+)诱导SH-SY5Y细胞建立PD细胞模型,通过Cell Counting Kit-8细胞毒性(CCK-8)实验确定MPP+的最佳作用浓度,并在成功建立的PD细胞模型中验证miR-1246的表达水平。利用TargetScan、miRWalk、miRase数据库对miR-1246的靶基因进行预测,并进行基因本体论(GO)富集分析和京都基因与基因组百科全书(KEGG)富集分析。结果PD组与对照组在年龄、性别上具有可比性。与对照组相比,PD组血清外泌体中miR-1246表达下调[0.73(0.20,1.21)vs.2.21(1.00,3.05),P<0.05]。ROC曲线分析显示miR-1246的曲线下面积为0.88,95%CI为0.77~0.99,截断值为0.98(最佳截断值)时的灵敏度为76.74%、特异度为95.00%。GO富集分析表明靶基因作用机制涉及细胞连接组装、对肽激素的细胞反应、γ-氨基丁酸突触、突触后膜、转录辅助激活蛋白结合方面,KEGG富集分析涉及癌症中的糖蛋白通路、磷脂酰肌醇3-激酶-蛋白激酶B(PI3K-AKT)信号通路、长寿调节途径、多巴胺能神经突触、肾素-血管紧张素系统(RAS)信号通路等。与正常SH-SY5Y细胞相比,在MPP+诱导的PD细胞模型中miR-1246的表达下调(2.16±1.69 vs.22.18±6.18,P<0.05)。结论miR-1246在PD患者血清外泌体及PD细胞模型中的表达下调,ROC曲线分析证实miR-1246在PD的诊断中具有预测能力,表明其或可作为PD的潜在生物标志物。Objective To investigate the diagnostic value of exosomal miR-1246 for Parkinson’s disease(PD).Methods Serum samples were collected from 30 PD patients(PD group)and 30 healthy controls(control group)receiving physical examination in the Second Affiliated Hospital of Xinjiang Medical University from September 2022 to April 2023.The expression level of miR-1246 in exosomes was analyzed using real-time quantitative PCR.The predictive value of miR-1246 in the diagnosis of PD was assessed using the receiver operating characteristic(ROC)curves.SH-SY5Y cells were induced by 1-methyl-4-phenylpyridinium(MPP+)to establish a PD cell model.The optimal concentration of MPP+was determined by CCK-8 assay,and the expression level of miR-1246 was verified in the MPP+-induced SH-SY5Y cell model.The target genes of miR-1246 were predicted using TargetScan,miRWalk,and miRase databases,and gene ontology(GO)analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)functional enrichment analysis were performed.Results Age and gender were matched between the PD and control groups.Compared with healthy controls,miR-1246 expression was significantly down-regulated in serum exosomes of PD patients[0.73(0.20,1.21)vs.2.21(1.00,3.05),P<0.05].ROC curve analysis showed that the area under the ROC curve(AUC)of miR-1246 was 0.88 with a 95%CI of 0.77-0.99.When the cutoff value was 0.98(optimum cutoff value),the sensitivity was calculated as 76.74%and the specificity was 95.00%.GO enrichment analysis indicated that the mechanism of action of target genes was involved with cellular junction assembly,cellular response to peptide hormones,GABA synapses,post-synaptic membranes,and transcriptional coactivator protein binding.KEGG signaling pathway was involved with glycoprotein pathway,PI3K-AKT signaling pathway,longevity regulation pathway,dopaminergic synapses,and RAS signaling pathway in cancer.In the MPP+-induced SH-SY5Y cell model,miR-1246 expression was significantly down-regulated compared with the control group(2.16±1.69 vs.22.18±6.18,P<0.05

关 键 词：帕金森病 血清外泌体 miR-1246 SH-SY5Y细胞 

分 类 号：R742.5[医药卫生—神经病学与精神病学]