摘要目的:探究重度抑郁障碍(MDD)患者mRNA中m6A甲基化修饰模式的特征及其在疾病发生中的作用机制。方法:2022年3月至2023年5月在哈尔滨市第一专科医院招募5例未接受过治疗的MDD患者(MDD组)和5名健康对照者(健康对照组)。通过微阵列分析确定MDD中mRNA的m6A修饰谱和基因表达模式。通过基因本体论(GO)富集分析和京都基因与基因组百科全书(KEGG)通路分析初步阐明mRNA的m6A甲基化在抑郁症发生发展中的作用。最后，通过RNA甲基化免疫沉淀结合定量PCR(MeRIP-qPCR)验证mRNA(GRM4、CAMKK2)的m6A甲基化水平。使用R语言4.2.0软件，对数据进行 t检验和Fisher确切概率法检验。 结果:MDD患者与健康对照组的m6A mRNA修饰存在明显差异。在MDD患者中，共有513个mRNA(180个高甲基化和333个低甲基化)存在m6A修饰差异。GO富集分析和KEGG通路分析表明，m6A高甲基化的mRNA主要参与神经活性配体与受体的相互作用，而m6A低甲基化的mRNA主要参与腺苷酸活化蛋白激酶(AMPK)信号通路。此外，共筛选出350个差异表达的mRNA，其中171个上调，179个下调，富集于环磷酸腺苷(cAMP)和肿瘤坏死因子(TNF)信号通路。MeRIP-PCR分析表明，MDD患者GRM4的m6A甲基化水平(25.40±2.38)高于健康对照组(9.40±1.00)( t＝13.88， P<0.05)，而CAMKK2的甲基化水平(19.63±6.60)则低于健康对照组(30.51±7.20)( t＝2.48， P<0.05)。 结论:m6A修饰表达谱在MDD患者中存在异常，其可能参与了MDD的发病和发展，而关键通路的鉴定可能为开发更有效的MDD治疗靶点提供新的线索和证据。

abstractsObjective:To investigate the characteristics of m6A methylation modification patterns in mRNA of patients with major depressive disorder (MDD) and its effect in the pathogenesis of the disease.Methods:From March 2022 to May 2023, five untreated MDD patients were assigned to the MDD group, and five healthy individuals were enrolled as the healthy control group at the First Psychiatric Hospital of Harbin.Microarray analysis was performed to determine the m6A modification profiles and gene expression patterns of mRNA in MDD. Gene ontology (GO) enrichment analysis and Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis were conducted to elucidate the effect of m6A methylation in the development of depression. Finally, methylated RNA immunoprecipitation combined with quantitative PCR (MeRIP-qPCR) was used to validate the m6A methylation levels of key mRNAs (GRM4, CAMKK2). Data were analyzed using R software (version 4.2.0) with t-test and Fisher's exact test. Results:Significant differences in m6A-modified mRNAs were observed between MDD patients and healthy controls. A total of 513 mRNAs (180 hypermethylated and 333 hypomethylated) exhibited differential m6A modifications in MDD patients. GO and KEGG analysis revealed that hypermethylated mRNAs were primarily enriched in neuroactive ligand-receptor interactions, while hypomethylated mRNAs were associated with the AMP-activated protein kinase (AMPK) signaling pathway. Additionally, a total of 350 differentially expressed mRNAs were identified (171 upregulated and 179 downregulated), enriched in the cyclic adenosine monophosphate (cAMP) and tumor necrosis factor (TNF) signaling pathways. MeRIP-qPCR results demonstrated that the m6A methylation level of GRM4 in MDD patients (25.40±2.38) was significantly higher than that in healthy controls (9.40±1.00) ( t=13.88, P<0.05), whereas the methylation level of CAMKK2 (19.63±6.60) was significantly lower than that in healthy controls (30.51±7.20) ( t=2.48, P<0.05). Conclusion:The m6A modification expression profile is abnormal in patients with major depressive disorder, which may be involved in the pathogenesis and development of MDD, and the identification of key pathways may provide new clues and evidence for the development of more effective therapeutic targets for MDD.