基于NMR技术对Wilson病模型大鼠尿液的代谢组学研究

1H-NMR Spectroscopy-based Metabonomic Research on Urine of Model Rats of Wilson＇s Disease

目的:以基于核磁共振(Nuclear Magnetic Resonance,NMR)的代谢组学方法对Wilson病(Wilson's Disease,WD)铜负荷模型大鼠及正常对照组大鼠的尿液进行研究,分析模型大鼠尿液中代谢物的变化,继而从小分子层面探讨铜过量对机体的损伤机制,以更加清楚的认识本病。方法:28只雄性Wistar大鼠,体重(180±20)g,随机被分为模型组(n=14)和健康对照组(n=14)。采用铜负荷法制作Wilson病大鼠模型,以NMR技术对大鼠尿液进行检测。采用MestRe-C 2.3软件及自编软件对谱图进行手动调相、基线校正和谱峰对齐。对样品进行分段积分,将积分数据归一化后构成数据矩阵,并利用PCA方法对数据矩阵进行统计分析。结果:相对于正常对照组,模型组大鼠尿液醋酸盐(acetate)含量有显著升高,柠檬酸盐(citrate)、苯乙酰甘氨酸(PAG)、琥珀酸盐(succinate)、甲胺(methylamine)、肌氨酸+肌氨酸酐(creatine/creatinine)、丙酮酸盐(pyruvate)、二甲基甘氨酸(DMG)、丙氨酸(alanine)含量有所升高,胆碱(choline)、牛磺酸(taurine)含量有所降低。这些发生改变的代谢物可能是潜在的WD铜负荷小分子代谢标志物,可为进一步研究WD的铜过量代谢机制提供参考。

Objective ： Applying IH nuclear magnetic resonance （ IH-NMR ） based metabonomics to study the changes of small molecular metabolites in the urine of the model rats of Wilson＇s disease. To explore the pathogenesis of Wilson＇s disease in small molecular aspect. Methods： 28 male Wistar rats[weight=（180 ± 20） g] were divided into two groups randomly, the model group （ n=14 ）and the control group （ n=14 ）, with the models established by copper-loaded method. Urine of the rats was tested with JH-NMR technology. The spectra was edited with MestRe-C2.3 and self-programmed software and then principal component analysis （ PCA ） was applied to differentiate the two groups. Results ： Acetate concentration was found to be significantly higher in the urine of the model group ; citrate, PAG, suceinate, methylamine, creatine/creatinine, pyruvate, DMG, and alanine were higher, and choline and taurine were lower in the urine of the model group. The small molecular metabolites mentioned above may contribute to the discrimination of the two groups, and provide references for further researches on the pathogenesis of WD.