Multiple sclerosis (MS) is a neurodegenerative disorder characterized by periodic neuronal demyelination, which leads to a range of symptoms and eventually to disability. The goal of this research was to use UPLC-Orbitrap/MS to identify validated biomarkers and explore the metabolic mechanisms of MS in mice. Thirty-two C57BL/6 male mice were randomized into two groups that were fed either normal food or 0.2% CPZ for 11 weeks. The mouse demyelination model was assessed by LFB and the expression of MBP by immunofluorescence and immunohistochemistry. The metabolites of the corpus callosum were quantified using UPLC-Orbitrap/MS. The mouse pole climbing experiment was used to assess coordination ability. Multivariate statistical analysis was adopted for screening differential metabolites, and the ingenuity pathway analysis (IPA) was used to reveal the metabolite interaction network. We successfully established the demyelination model. The CPZ group slowly lost weight and showed an increased pole climbing time during feeding compared to the CON group. A total of 81 metabolites ( and ) were determined to be enriched in 24 metabolic pathways; 41 metabolites were markedly increased, while 40 metabolites were markedly decreased in the CPZ group. The IPA results revealed that these 81 biomarker metabolites were associated with neuregulin signaling, PI3K-AKT signaling, mTOR signaling, and ERK/MAPK signaling. KEGG pathway analysis showed that two significantly different metabolic pathways were enriched, namely, the glycerophospholipid and sphingolipid metabolic pathways, comprising a total of nine biomarkers. Receiver operating characteristic analysis showed that the metabolites (e.g., PE (16 : 0/22 : 6(4Z, 7Z, 10Z, 13Z, 16Z, 19Z)), PC (18 : 0/22 : 4(7Z, 10Z, 13Z, 16Z)), cytidine 5-diphosphocholine, PS (18 : 0/22 : 6(4Z, 7Z, 10Z, 13Z, 16Z, 19Z)), glycerol 3-phosphate, SM (d18 : 0/16 : 1(9Z)), Cer (d18:1/18 : 0), galabiosylceramide (d18:1/18 : 0), and GlcCer (d18:1/18 : 0)) have good discrimination ability for the CPZ group. In conclusion, the differential metabolites have great potential to serve as biomarkers of demyelinating diseases. In addition, we identified metabolic pathways associated with CPZ-induced demyelination pathogenesis, which provided a new perspective for understanding the relationship between metabolites and CNS demyelination pathogenesis.

中文翻译：

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通过 UPLC-Orbitrap/MS 对 Cuprizone 诱导的小鼠胼胝体脱髓鞘的非靶向代谢组学分析揭示了 CNS 脱髓鞘疾病的潜在代谢生物标志物

多发性硬化症 (MS) 是一种以周期性神经元脱髓鞘为特征的神经退行性疾病，可导致一系列症状并最终导致残疾。本研究的目的是使用 UPLC-Orbitrap/MS 来识别经过验证的生物标志物并探索小鼠 MS 的代谢机制。将 32 只 C57BL/6 雄性小鼠随机分为两组，分别喂食正常食物或 0.2% CPZ 11 周。通过LFB评估小鼠脱髓鞘模型，并通过免疫荧光和免疫组织化学评估MBP的表达。使用 UPLC-Orbitrap/MS 对胼胝体的代谢物进行量化。鼠标爬杆实验用于评估协调能力。采用多元统计分析筛选差异代谢物，并使用独创性途径分析（IPA）揭示代谢物相互作用网络。我们成功建立了脱髓鞘模型。与 CON 组相比，CPZ 组体重缓慢下降，并且在喂食期间显示出增加的爬杆时间。共有 81 种代谢物 (和)被确定富含 24 种代谢途径；CPZ组有41种代谢物显着增加，而40种代谢物显着减少。IPA 结果显示，这 81 种生物标志物代谢物与神经调节蛋白信号、PI3K-AKT 信号、mTOR 信号和 ERK/MAPK 信号相关。KEGG通路分析显示，富集了两条明显不同的代谢途径，即甘油磷脂和鞘脂代谢途径，共包含9个生物标志物。接受者操作特征分析表明代谢物（例如，PE（16：0/22：6（4Z、7Z、10Z、13Z、16Z、19Z））、PC（18：0/22：4（7Z、10Z、13Z） , 16Z)), 胞苷 5 -二磷酸胆碱，PS（18：0/22：6（4Z，7Z，10Z，13Z，16Z，19Z）），3-磷酸甘油，SM（d18：0/16：1（9Z）），Cer（d18：1 /18:0)、galabosylceramide (d18:1/18:0)、GlcCer (d18:1/18:0))对CPZ组有良好的辨别能力。总之，差异代谢物具有作为脱髓鞘疾病生物标志物的巨大潜力。此外，我们确定了与CPZ诱导的脱髓鞘发病机制相关的代谢途径，这为理解代谢物与CNS脱髓鞘发病机制之间的关系提供了新的视角。