Equine and Companion Animal Nutrition, Department of Morphology, Imaging, Orthopedics, Rehabilitation and Nutrition, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820, Merelbeke, Belgium.
Small Animal Department, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820, Merelbeke, Belgium.
Sci Rep. 2024 Jun 26;14(1):14750. doi: 10.1038/s41598-024-64777-z.
Cerebrospinal fluid metabolomics is a promising research technology in the elucidation of nervous system disorders. Therefore, in this work, a cerebrospinal fluid (CSF) metabolomics method using liquid chromatography coupled to mass spectrometry was optimized and validated to cover a wide range of metabolites. An acceptable coefficient of variance regarding instrumental, within-lab and intra-assay precision was found for 95, 70 and 96 of 102 targeted metabolites, together with 1256, 676 and 976 untargeted compounds, respectively. Moreover, approximately 75% of targeted metabolites and 50% of untargeted compounds displayed good linearity across different dilution ranges. Consequently, metabolic alterations in CSF of dogs with idiopathic epilepsy (IE) were studied by comparing CSF of dogs diagnosed with IE (Tier II) to dogs with non-brain related disease. Targeted metabolome analysis revealed higher levels of cortisol, creatinine, glucose, hippuric acid, mannose, pantothenol, and 2-phenylethylamine (P values < 0.05) in CSF of dogs with IE, whereas CSF of dogs with IE showed lower levels of spermidine (P value = 0.02). Untargeted CSF metabolic fingerprints discriminated dogs with IE from dogs with non-brain related disease using Orthogonal Partial Least Squares Discriminant Analysis (R(Y) = 0.997, Q(Y) = 0.828), from which norepinephrine was putatively identified as an important discriminative metabolite.
脑脊液代谢组学是阐明神经系统疾病的一种很有前途的研究技术。因此，在这项工作中，我们优化和验证了一种基于液相色谱-质谱联用的脑脊液代谢组学方法，以覆盖广泛的代谢物。对于 102 种靶向代谢物中的 95、70 和 96 种，仪器、实验室内部和测定内精密度的变异系数分别可接受，对于 1256、676 和 976 种非靶向化合物也分别可接受。此外，大约 75%的靶向代谢物和 50%的非靶向化合物在不同稀释范围内显示出良好的线性关系。因此，通过比较诊断为特发性癫痫（IE）的犬的脑脊液与无脑部疾病犬的脑脊液，研究了犬 IE 脑脊液中的代谢变化。靶向代谢组学分析显示，IE 犬脑脊液中皮质醇、肌酸、葡萄糖、马尿酸、甘露糖、泛醇和 2-苯乙胺（P 值均<0.05）水平较高，而 IE 犬脑脊液中 spermidine（P 值=0.02）水平较低。基于正交偏最小二乘判别分析（R(Y)=0.997，Q(Y)=0.828），可通过非靶向 CSF 代谢指纹图谱将 IE 犬与无脑部疾病犬区分开来，其中去甲肾上腺素被推测为一种重要的鉴别代谢物。
