肝气郁结证大鼠尿液代谢组学研究

Study on urine metabonomics for rat model with stagnation of liver-Qi syndrome

目的:探讨肝气郁结证对大鼠体内尿液小分子内源性代谢物的影响,以及造模前后机体内代谢通道的变化,从整体的角度来阐述肝气郁结证的生物学本质。方法:20只SD大鼠,体重(240±28)g,随机分为两组:正常组10只(A组)、模型组10只(B组),采用夹尾法造模建立大鼠肝气郁结证模型。分别在造模前1天、成模后收集尿液,通过NMR波谱仪检测,运用代谢组学的方法,观察与分析两组大鼠尿液小分子代谢物组的差异性变化谱。结果:与正常组相比,模型组大鼠尿液马尿酸3.97、7.55、7.85,α-酮戊二酸3.01、2.45,柠檬酸(citrate)4.10、2.88、2.57,异柠檬酸(isocitrate)3.99、2.51、2.43和乌头酸(aconitate)3.77、6.96、3.96的含量降低,肌酸酐3.05、4.07,丙酮2.23,乙酸1.93,肌酸3.93、3.03,烟酸(nicotinate)4.41、1.41和5-羟(基)吲哚-3-乙酸(5-hydroxyindole-3-acetate,5-HIAA)3.03、2.91、6.63、7.15谱峰相对积分面积明显增高,在尿液中的含量显著升高。结论:运用代谢组学尿液研究方法可以成功区分肝气郁结证模型组与正常组大鼠的尿液,初步得到其证型的代谢表型与差异性代谢物。尿液代谢组学的变化可以反映大鼠的生理病理基础,阐述肝气郁结证的生物学本质。

Objective：To investigate the influence of stagnation of liver-Qi syndrome（SLQS） on endogenous small-molecule metabolites in urine of rat model and the changes of metabolism channel before and after modeling,in order to expatiate the biological foundation of SLQS.Methods：Twenty healthy SD rats,body weight （240±28） gramme,were randomly divided into two groups：One was normal controlled group（group A）,had 10 rats,fed by routine,the other was experimental group（group B） made into model of SLQS by nipping cauda.The urine samples of two group rats were collected on the day before modeling and the tenth day after modeling,and treated by NMR spectrometer and principal component analysis.Results：Comparing with the rats’ urine of group A,the urine contents of hippuric acid 3.97,7.55,7.85,alpha-ketoglutarate 3.01,2.45,citrate 4.10,2.88,2.57,isocitrate 3.99,2.51,2.45,and aconitate 3.77,6.96,3.96 were decrease in group B;the contents and curve relative integral acreage of creatinine 3.05,4.07,acetone 2.23,acetic acid 1.93,reatine 3.93,3.03,nicotinate 4.41,1.41,and acid 5-hydroxyindole-3-acetate（5-HIAA） 3.03,2.91,6.63,7.15 were increase in group B.Conclusion：Using the methods of metabonomics can succesfully identify the rats’ urine of group A and B,primarily gain the metabolic phenotype and diversity metabolites of rats.The metabonomics changes of rats’ urine can reflect the physiological and pathological basis of rat model with SLQS,expatiate its biological foundation.