摘要
目的分析阿尔茨海默病(AD)模型小鼠前额叶皮层的差异表达基因,从转录组水平揭示AD的发病机制。方法本研究随机选取9月龄雌性APP swe/PS1ΔE9(PAP)的模型小鼠和野生型C57BL/6 J小鼠各5只,提取前额叶皮层组织RNA,用Illumina HiSeq 3000测序。采用edgeR软件进行表达差异显著性分析。分析AD组和对照组基因表达变化,并采用qRT-PCR对其中6个关键差异表达基因进行验证。然后对差异表达基因进行聚类分析、GO功能富集分析、KEGG通路富集分析。结果在AD组与对照组之间发现224个差异表达基因(P<0.05,|logFC|> 1.0),其中205个基因上调,19个基因下调。6个关键基因qRT-PCR验证结果与RNA-Seq趋势一致。GO功能富集分析结果表明,这些差异表达基因与免疫反应、炎症反应、趋化因子活动以及IgG结合等有关;KEGG通路富集分析结果表明,这些基因参与吞噬、溶酶体、Toll样受体信号通路、细胞因子受体相互作用、NF-κB信号通路等重要生物学通路。结论得到AD相关差异表达基因,为利用模型小鼠进行AD相关机制和治疗研究提供了实验依据。
Objective To determine the transcriptomic profile of AD mice and controls by RNA-Seq,in order to identify differentially expressed genes and reveal the molecular mechanism of AD.Methods Nine-month-old APP/PS1 mice and age-matched wild-type C57BL/6 J mice were selected,with five female mice included in each group.Genome-wide analysis of mRNA expression in the prefrontal cortex of AD mice and controls was performed by RNA-Seq.qRT-PCR was used to verify the six key genes.Then,cluster analysis,Gene Ontology(GO)function enrichment analysis,and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were used to analyze gene function.Results The result showed that a total of 224 genes were differentially expressed in the two groups(P<0.05,|logFC|>1.0):205 genes were upregulated and 19 genes were downregulated.When qRT-PCR was used to validate the six key genes,the result were consistent with the RNA-Seq.GO enrichment analysis result showed that the differentially expressed genes are involved in immune response,inflammatory response,chemokine activity,and IgG binding.KEGG pathway enrichment analysis revealed that these genes participate in phagosome,lysosome,toll-like receptor signaling pathway,cytokine-cytokine receptor interactions,and the NF-κB signaling pathway.Conclusions Differentially expressed genes related to AD were obtained,which provides an important experimental basis for the usage of a double-transgenic mouse model to study AD-related mechanisms and pharmacological intervention.
作者
史长华
张玲
陈巍
付信靖
秦川
SHI Changhua;ZHANG Ling;CHEN Wei;FU Xinjing;QIN Chuan(Comparative Medicine Center,Peking Union Medical College(PUMC);Institute of Laboratory Animal Science, Chinese Academy of Medical Science(CAMS);Key Laboratory of Human Disease Comparative Medicine, Ministry of Health,Beijing 100021,China)
出处
《中国比较医学杂志》
CAS
北大核心
2018年第10期1-7,共7页
Chinese Journal of Comparative Medicine
基金
国家自然基金(81500938)
科技部国合项目(2015DFG32230)。