【摘要】 目的筛选氧化应激介导的阿尔茨海默病(AD)相关的microRNAs(miRNAs),并进行验证。方法利用基因芯片分别筛选体外培养的原代海马神经元与氧化应激海马神经元及正常老化小鼠(SAMR1)与速老化小鼠(SAMP8)海马组织差异miRNAs,两组芯片结果取交集后获得共同差异表达miRNAs。进一步对两组芯片共表达差异miRNAs进行Real-time PCR验证和生物信息学分析。结果 miRNA芯片结果显示,与体外培养的原代海马神经元相比,氧化应激的海马神经元中101个miRNAs表达改变(64个表达上调,37个表达下调)。与SAMR1鼠比,SAMP8海马组织中294个miRNAs表达改变(131个表达上调,163个表达下调)。其中有6个差异表达的miRNAs (miR-296、miR-20a、miR-329、miR-193b、miR-130b和miR-24)在体外氧化应激的海马神经元和SAMP8鼠海马组织中表达的miRNAs中表达均上调,而miR-376b则在两种模型中表达均下调。应用Real-time PCR对上述两种模型中共同表达趋势一致的miRNAs进行验证,结果与芯片检测结果一致。进一步对两组芯片共表达的7个差异miRNAs进行生物信息学分析,结果显示,差异miRNAs的靶基因共调控1 443个靶基因。这些靶基因主要参与神经发育、MAPK信号通路和内吞作用等。结论氧化应激可介导miR-296、miR-20a、miR-329、miR-193b、miR-130b、miR-24和miR-376b表达失调,可能通过影响神经发育、MAPK信号通路和内吞作用等参与AD的发生发展。更多还原

【Abstract】 Objective To screen and validate the oxidative stress-mediated microRNAs(miRNAs) associated with Alzheimer’s disease(AD) that could be used as diagnostic markers and research targets or for AD. Methods The expression of miRNAs in primary hippocampal neurons and oxidative stressed primary hippocampal neurons, as well as in the hippocampus tissue of SAMR1 and SAMP8 mice were detected by microarray. The results of two sets of microarray were combined to screen out the co-expressed differential miRNAs. Furthermore, the co-expressed differential miRNAs were validated by the real time PCR and bioinformatics analysis was carried out for their function prediction.Results Gene chip results showed that the expression of 101 microRNAs in oxidative stressed primary hippocampal neurons were altered compared with that in primary hippocampal neurons(64 were up-regulated and 37 were down-regulated). Compared with SAMR1 mice, there were 294 miRNAs expression changes in hippocampus of SAMP8 mice(131 miRNAs were up-regulated and 163 miRNAs were down-regulated). Six of co-expressed differential miRNAs(miR-296, miR-20 a, miR-329, miR-193 b, miR-130 b, and miR-24) in the two sets of microarray were up-regulated, while miR-376 b was down-regulated in both sets. The results of Real-time PCR were consistent with the results of microarray analysis, and bioinformatics analysis showed that 1 443 target genes were regulated by co-expressed differential miRNAs. These target genes were mainly involved in neurodevelopment, MAPK signaling pathway and endocytosis.Conclusions Oxidative stress could mediate the dysregulation of miR-296, miR-20 a, miR-329, miR-193 b, miR-130 b, miR-24, and miR-376 b, and involve in the development of AD possibly through affecting axon guidance, MAPK signaling pathway signaling and endocytosis, which provides an experimental basis for further exploring the mechanism of oxidative stress in the pathogenesis of AD.更多还原