阿尔茨海默病中高表达的miR-182-5p靶向抑制了Neuritin的表达  

Up-regulated miR-182-5p targeting inhibits the expression of Neuritin in Alzheimer′s disease

作　　者：孙嘉伟 宋银 宋丹丹 张雪[1] 黄娟 章云衡 黄瑾[1] SUN Jiawei;SONG Yin;SONG Dandan;ZHANG Xue;HUANG Juan;ZHANG Yunheng;HUANG Jin(Medical College of Shihezi University,Shihezi,Xinjiang 832000,China;Department of medicine,Hangzhou Normal University,Hangzhou,Zhejiang 310036,China)

机构地区：[1]石河子大学医学院,新疆石河子832000 [2]杭州师范大学医学部,浙江杭州310036

出　　处：《石河子大学学报（自然科学版）》2021年第4期503-510,共8页Journal of Shihezi University(Natural Science)

基　　金：国家自然科学基金面上项目(8177051901);国家科技重大专项(民口)重大新药创制项目(2019ZX09301-161)。

摘　　要：目的筛选阿尔茨海默病(Alzheimer′s disease,AD)数据集中可能靶向Neuritin的目标miRNA,明确二者在AD动物模型中的相关性,进一步探讨目标miRNA对Neuritin的靶向抑制作用。方法从AD小鼠脑组织miRNA数据集中,富集AD中上调的miRNA;同时利用生物信息学手段,分析预测靶向Neuritin的miRNA;二者取交集,筛选获得AD中上调的可能靶向Neuritin的目标miRNA。为了明确二者的相关性,我们在AD模型小鼠中,使用qRT-PCR检测目标miRNA在脑组织的表达水平,同时利用Western Blot检测Neuritin蛋白在脑组织的表达。然后,在细胞水平观察了目标miRNA模拟物对Neuritin表达的影响。最后,我们用荧光素酶报告基因实验探讨了目标miRNA对Neuritn 3′UTR区特异性的结合作用。结果筛选获得AD中上调、且可能靶向Neuritin的目标miRNA———miR-182-5p;在AD模型小鼠海马组织中,miR-182-5p的表达升高(P<0.05),且Neuritin的表达降低(P<0.01),说明AD模型小鼠海马中miR-182-5p与Neuritin的表达呈负相关;miR-182-5p模拟物转染293T细胞后,Neuritin表达低于对照组(P<0.05),说明miR-182-5p抑制了Neuritin的表达;荧光素酶报告基因结果显示,共转染荧光素酶报告基因和miR-182-5p模拟物后,pLUC-Neuritin组的荧光活性低于pLUC-MUT-Neuritin组(P<0.01),说明miR-182-5p通过与Neuritn 3′UTR区特异结合,靶向抑制了Neuritin的表达。结论AD模型小鼠中miR-182-5p表达升高,miR-182-5p可以靶向抑制Neuritin的表达。Objective To screen the miRNA targeting Neuritin in Alzheimer′s disease(AD)datasets,to clarify their correlation and explore the inhibition effect of the target miRNA on Neuritin in AD model mice.Method We enriched the up-regulated miRNAs from AD mice brain miRNA datasets,and predicted the miRNAs targeting Neuritin by using bioinformatics methods.In order to clarify the correlation,we detected the expression of target miRNA and Neuritin protein by qRT-PCR and Western blot in the brain of AD mice.Then,the effect of target miRNA mimics on Neuritin expression was observed at the cellular level.Finally,we used luciferase reporter gene experiment to explore the targeted regulation of target miRNA on Neuritn 3′UTR.Result miR-182-5p was selected as the target miRNA of Neuritin which may up-regulated in AD.In hippocampus of AD model mice the expression of miR-182-5p was increased(P<0.05),and the expression of Neuritin was decreased(P<0.01),which indicated the negative correlation between miR-182-5p and Neuritin.The expression of Neuritin in 293T cells transfected with miR-182-5p mimics was lower than that in control group(P<0.05),indicating that miR-182-5p inhibited the expression of Neuritin.Luciferase reporter gene results showed that the fluorescence activity of pLUC-Neuritin group was lower than that of pLUC-MUT-Neuritin group(P<0.01),indicating that miR-182-5p inhibits the expression of Neuritin by binding to 3′UTR region.Conclusion The expression of miR-182-5p was increased in AD model mice,and miR-182-5p could target inhibit the expression of Neuritin.

关 键 词：miR-182-5p NEURITIN 阿尔茨海默病 生物信息学 

分 类 号：R749.16[医药卫生—神经病学与精神病学]