儿童哮喘患者尿液中小分子代谢物分析

Analysis on Small Molecule Metabolites in Urine of Patients with Childhood Asthma

目的采用基于气相色谱质谱联用(GC/MS)的代谢组学技术研究儿童哮喘患者尿液中小分子代谢物的变化。方法取50例儿童哮喘患者尿样及49例年龄、性别匹配的正常儿童尿样,用GC/MS技术检测尿液中的小分子代谢物,应用正交偏最小二乘法(OPLSA)的方法,观察正常儿童对照组和儿童哮喘组的代谢轮廓差异。结果正常儿童对照组和儿童哮喘组的代谢谱能明显的区分,利用商业化的代谢物谱库(如Wiley和NIST质谱库)及本实验室建立的标准品代谢物谱库鉴定了18个差异代谢物。与正常儿童对照组比较,儿童哮喘患者磷酸盐、L-苏氨酸含量降低;丁二酸、L-脯氨酸、β-D-呋喃木糖、肌酐、2,3-二羟基丁酸、苯乙酸、D-阿拉伯糖、1H-吲哚、木糖醇、α-L-吡喃甘露糖、苯乙酸、α-L-吡喃甘露糖、甘氨酸、D-葡萄糖、肌醇、尿酸、马尿酸、1H-吲哚-3-乙酸含量升高。结合代谢途径分析发现,儿童哮喘患者体内存在能量代谢、肠道菌群代谢、氨基酸代谢等代谢异常。结论基于GC/MS的代谢组学技术能全面客观地反映儿童哮喘患者体内尿液中的代谢物变化,潜在的代谢标志物分析及代谢途径探讨将为儿童哮喘的诊断和发病机制研究提供一个可借鉴的思路和手段。

Abstract Objective To investigate the characteristics of urine metabolites in children with asthma. Methods 50 patients of children with asthma and 49 age, sex matched volunteers were enrolled and their urine samples were collected with GC/MS. Orthogonal--Projection to Latent Structures （OPLS） were performed to detect the metabolic profile difference between the children with asthma and control group. Results Children with asthma can be precisely distinguished from normal children in the control group. 18 differences in metabolites were identified using commercial metabolite library （such as Wiley and NIST mass spectral library） and the establishment of the laboratory＇s standard library metabolites. Compared with normal children in the control group, levels of phosphate, L- threonine were decreased; while the levels of succinic acid, L- proline, β -D - xylofuranose, creatinine, 2,3 -hydroxybutyric acid, phenylacetic acid, D - arabinose, 1H - indole, xylitol, α - L - mannopyranosyl, phenylacetic acid, α - L - mannopyranosyl, glycine, D - glucose, inositol, uric acid, hippuric acid, 1H - indole - 3 - acetic acid were increased, in children with asthma group. With anaIysis of metabolic pathway, the dysfunction of energy metabolism, intestinal flora metabolism, amino acid metabolism and other metabolic abnormalities were found in vivo with children with asthma. Conclusion Work shows that metabonomics based on GS/MS can be used to comprehensively and objectively reflect the changes in the body of metabolites in the urine of children with asthma. The analysis of underlying metabolic markers and metabolic pathway will provide a draw ideas and methods for clinical diagnosis and pathogenesis of asthma study.