APP/PS1双转基因阿尔茨海默病小鼠海马组织环状RNA N6-甲基腺苷甲基化修饰的初步研究  

Preliminary study on circRNA N6-methyladenosine(m6A)methylation in hippocampus of APP/PS1 double transgenic mice with Alzheimer's disease

作　　者：闫林娜 熊静 吕文静 张鑫[1] 王梓炫[1] Yan Linna;Xiong Jing;Lyv Wenjing;Zhang Xin;Wang Zixuan(Department of Geriatrics,the Affiliated Hospital of Qingdao University,Qingdao 266000,China)

机构地区：[1]青岛大学附属医院老年医学科,青岛266000

出　　处：《中华老年医学杂志》2025年第2期188-193,共6页Chinese Journal of Geriatrics

基　　金：中国博士后科学基金面上资助(2019M662305);青岛市博士后应用研究项目;齐鲁卫生与健康领军人才培育工程专项经费资助。

摘　　要：目的分析阿尔茨海默病(AD)小鼠与野生型(WT)小鼠海马组织中环状RNA(circRNA)表达与N6-甲基腺苷(m6A)甲基化修饰水平,初步探究环状circRNA m6A甲基化修饰参与AD的机制。方法通过RNA高通量测序(RNA-seq)筛选APPswe/PS1dE9(APP/PS1)双转基因AD模型小鼠(3只)与相匹配的野生型小鼠(3只)海马组织中差异表达的circRNAs,并对其来源基因进行基因本体论(GO)和京都基因和基因组百科全书(KEGG)富集分析,实时荧光定量PCR(qRT-PCR)验证测序结果。通过SRAMP网站预测差异表达的circRNAs潜在的m6A修饰位点,利用甲基化RNA免疫共沉淀-定量PCR(MeRIP-qPCR)对具备很高可信度的m6A修饰位点进行验证分析。结果与对照组比较,APP/PS1组有21个显著差异表达的circRNAs,其中15个circRNAs上调,6个circRNAs下调。GO富集分析显示,差异表达circRNA的来源基因与突触结构和功能相关;KEGG富集分析显示,来源基因与Hippo通路、脂类代谢等通路有关。qRT-PCR结果显示circRNA表达趋势与测序结果一致。APP/PS1组mmu-Bai3_0007低于对照组(0.033±0.002比1.006±0.136,t=12.38,P<0.01),mmu-Bai3_0007 m6A甲基化修饰水平也低于对照组(0.144±0.018比0.431±0.087,t=5.62,P<0.01)。结论Bai3circRNA m6A甲基化修饰可能通过调控Bai3circRNA的表达来参与AD发病。Objective To assess the levels of circRNA and N6-methyladenosine(m6A)methylation in hippocampal tissue of Alzheimer's disease(AD)mice and wild-type(WT)mice,and to explore the potential role of circRNA m6A methylation modification in the pathogenesis of Alzheimer's disease.Methods Differentially expressed circular RNAs(circRNAs)in the hippocampal tissue of APPswe/PS1dE9(APP/PS1)double-transgenic Alzheimer's disease(AD)model mice and matched wild type mice were identified using RNA high-throughput sequencing(RNA-seq).Subsequently,Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were conducted on the source genes of these circRNAs.Validation of the RNA-seq results was performed using real-time quantitative PCR(qRT-PCR).Furthermore,the SRAMP website was utilized to predict potential m6A modification sites on the differentially expressed circRNAs.Finally,circRNAs with high confidence were validated and analyzed through methylated RNA immunoprecipitation-quantitative PCR(MeRIP-qPCR).Results In comparison to the control group,21 circRNAs exhibited differential expression in the APP/PS1 group,with 15 being up-regulated and 6 down-regulated.GO analysis revealed that the source genes of these differentially expressed circRNAs were primarily associated with synapse structure and function,while KEGG analysis indicated their involvement in the Hippo signaling pathway and Lipid metabolism.qRT-PCR results confirmed that the expression patterns of the differentially expressed circRNAs were consistent with the RNA-seq findings.Specifically,the expression of mmu-Bai3_0007 in the APP/PS1 group was significantly lower than that in the control group(0.033±0.002 vs.1.006±0.136,t=12.38,P<0.01),and the m6A methylation level of mmu-Bai3_0007 was also notably reduced in the APP/PS1 group compared to the control group(0.144±0.018 vs.0.431±0.087,t=5.62,P<0.01).Conclusions The m6A methylation modification of Bai3circRNA could play a role in Alzheimer's disease by regulating the expression of Bai3 circ

关 键 词：阿尔茨海默病 N6-甲基腺苷甲基化 环状RNA 

分 类 号：R749.16[医药卫生—神经病学与精神病学]