脾气虚证模型大鼠尿液的1H-NMR代谢组学研究  

~1H-NMR-based metabonomics study on urine of rat with Spleen-Qi deficiency pattern

作　　者：罗良 陈嘉辉 王园园 张晓君[1] 尹西拳 卢碧钰 李媛 郑海慧 谢智勇[3] 廖琼峰[1] 

机构地区：[1]广州中医药大学中药学院,广东广州510006 [2]无限极(中国)有限公司,广东广州510623 [3]中山大学药学院,广东广州510006

出　　处：《中国药理学通报》2017年第10期1363-1370,共8页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81473540,81473319);广东省科技计划项目(No 2014A020221027,2013B090800011,2014B040404010,2015A030401031);广东省自然科学基金资助项目(No 015A030313123)

摘　　要：目的建立脾气虚证大鼠模型,分析脾气虚证的代谢通路,探究内源性代谢产物变化与脾气虚证的关系。方法采用苦寒泻下、劳倦过度和饥饱失常复合因素方法建立并评价脾气虚证大鼠模型,检测其血清中肌酸磷酸激酶活性;对大鼠尿液进行~1H-NMR检测,以变化倍数(fold change,FC)>1.2,结合独立样本t检验(P<0.05)的统计学方法筛选脾气虚证模型组和对照组的差异代谢物,进行代谢通路分析与富集分析。结果脾气虚证模型建立成功;模型组的肌酸磷酸激酶活性低于对照组(P<0.05);从主成分分析(PCA)结果得出模型组和对照组的代谢产物得到明显区分;筛选出33种差异代谢物,主要参与的代谢通路涉及能量代谢、氨基酸代谢、核苷酸代谢,同时对肠道菌群存在干扰。结论脾气虚主要扰乱了大鼠的能量代谢通路(三羧酸循环、糖酵解、脂质氧化),抑制了供能作用,导致大鼠出现疲乏和体质量增长抑制的症状。Aim To establish the rat model of SpleenQi deficiency,analyse the metabolic pathways and investigate the connection between the changed urinary metabolites and Spleen-Qi deficiency,in order to explore the potential mechanisms of Spleen-Qi deficiency. Methods With the binding methods of diarrhea induced by bitter and cold,abnormal of starvation and excessive tiredness,the rat Spleen-Qi deficiency model was established. Then the activity of creatine phosphokinase（ CPK） was detected. The endogenous metabolites in the urine were detected by NMR,and the data were analyzed with multivariate and statistical methods. Then the metabolites were selected that could be clearly distinct in the two groups with the fold change value（ 1. 2） and the P 0. 05 of Student＇s ttest. Both the pathway analysis and enrichment analysis were performed with Metabo Analyst 3. 0. Results Compared with the normal rats,the activity of CPK decreased significantly in model rats（ P 0. 05）. A significant separation appeared in the principal components analysis（ PCA） score plot when the control group and the model group were compared,indicating that the Spleen-Qi deficiency model was successfully duplicated. The 33 differential metabolites,which mainly involved in the metabolic pathways,were distinguished from the comparision of Spleen-Qi deficiency model group and control group. The metabolic pathways was related to energy metabolism,amino acid metabolism,nucleotide metabolism and disturbance of gut microbes. Conclusions The main energy metabolic pathways（ tricarboxylic acid cycle,glycolysis and liquid oxidation） may be disturbed in Spleen-Qi deficiency rats. The energy supply function is suppressed,which leads to the fatigue and weight loss in rats.

关 键 词：脾气虚证 代谢组学 尿液 核磁共振 内源性代谢物 代谢通路 

分 类 号：R-332[医药卫生] R256.39