阿尔茨海默病血清外泌体miRNA生物标志物的筛选研究  

Screening of serum exosomal miRNAs as diagnostic biomarkers in Alzheimer’s disease

作　　者：段贤 郑庆[1] 梁力晖[1] 周琳[2,3] DUAN Xian;ZHENG Qing;LIANG Lihui;ZHOU Lin(Department of Geriatrics,Hunan Provincial People’s Hospital,Changsha,Hunan Province,410002;Department of Geriatric Neurology,Xiangya Hospital,Central South University,Changsha,Hunan Province,410008;Department of Neurology,Second Affiliated Hospital of Xinjiang Medical University,Urumqi,Xinjiang Uygur Autonomous Region,830063,China)

机构地区：[1]湖南省人民医院老年医学科,长沙410002 [2]中南大学湘雅医院老年神经内科,长沙410008 [3]新疆医科大学第二附属医院神经内科,乌鲁木齐830063

出　　处：《陆军军医大学学报》2024年第15期1803-1810,共8页Journal of Army Medical University

基　　金：新疆神经系统疾病研究重点实验室开放课题(XJDX1711-2214)。

摘　　要：目的通过比较阿尔茨海默病(Alzheimer’s disease,AD)患者与健康对照者的血清外泌体中miRNAs的表达,筛选出差异表达的miRNAs。方法收集2017年3月至2018年8月就诊于中南大学湘雅医院老年神经内科的AD患者71例为研究组(AD),健康体检者71例为对照组(HC)。各组选取4例样本行血清外泌体miRNAs高通量二代测序,检测、分析2组间差异表达的miRNAs,筛选出表达差异有统计学意义、表达量最高的前4种miRNAs,在71例AD患者和71例健康体检者样本中用实时荧光定量逆转录聚合酶链式反应(qRT-PCR)测定其相对浓度。结果高通量二代测序共检测到775种miRNAs,其中有44种miRNAs在2组间表达差异有统计学意义(P<0.05)。与HC组相比,AD组中有34种miRNAs表达上调,10种miRNAs表达下调,其中表达量最高的前4种miRNAs为miRNA-148a-3p、miRNA-16-5p、miRNA-19b-3p、miRNA-483-5p。MiRNA-148a-3p表达在AD组中较HC组显著上调(P<0.01),但miRNA-16-5p、miRNA-19b-3p、miRNA-483-5p在2组间表达差异无统计学意义。绘制ROC曲线,miRNA-148a-3p的AUC为0.7113(95%CI:0.622~0.801),灵感度为71.6%,特异度为69.7%。结论血清外泌体miRNA-148a-3p可望作为AD诊断的生物标志物。Objective To screen differentially expressed miRNAs(DEMs)by comparing the expression of miRNAs in serum exosomes between Alzheimer’s disease(AD)patients and healthy controls.Methods A total of 71 AD patients admitted to Department of Geriatric Neurology of Xiangya Hospital from March 2017 to August 2018 and another 71 healthy individuals who taking physical examination in the hospital during same period were recruited and assigned into AD and HC groups,respectively.Four AD patients and four healthy subjects were selected for high-throughput second-generation sequencing of exosome miRNAs.The results were analyzed to obtain the DEMs between them,and the top 4 DEMs were finally selected.Then real-time quantitative real-time PCR was applied for all the subjects to detect the expression of the 4 DEMs.Results High-throughput second-generation sequencing detected 775 miRNAs,and 44 DEMs were found with statistical difference between the 2 groups(P<0.05).Compared with the HC group,34 miRNAs were up-regulated and 10 were down-regulated in the AD group.The top 4 DEMs were miRNA-148a-3p,miRNA-16-5p,miRNA-19b-3p and miRNA-483-5p.MiRNA-148a-3p was significantly up-regulated in the AD group than the HC group(P<0.01),but there were no significant differences in the expression level in the other 3 DEMs between the 2 groups.ROC curve analysis showed that the area under the curve of miRNA-148a-3p was 0.7113(95%CI:0.622~0.801),with a sensitivity of 71.6%and a specificity of 69.7%.Conclusion Serum exosome miRNA-148a-3p can be used as a biomarker for the diagnosis of AD.

关 键 词：阿尔茨海默病 外泌体 MIRNAS 生物标志物 

分 类 号：Q522[生物学—生物化学] R446.119[医药卫生—诊断学] R745.7[医药卫生—临床医学]