中国中药杂志

2017, v.42(24) 4855-4863

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脾气虚证大鼠尿液生物标记物的识别及其生物学意义
Identification of biomarkers in urine of rats with spleen Qi deficiency and biological significance

刘武平;李婵艺;黄晶;廖婧竹;马文杰;陈宏远;芮雯;
LIU Wu-ping;LI Chan-yi;HUANG Jing;LIAO Jing-zhu;MA Wen-jie;CHEN Hong-yuan;RUI Wen;Central Laboratory,Guangdong Pharmaceutical University;Department of Immunology & Microbiology,Guangdong Pharmaceutical University;

摘要(Abstract):

考察脾气虚证大鼠尿液小分子代谢组成分,寻找脾气虚证的标记物,阐明其与代谢通路之间的联系。采用劳倦伤脾加限制饮食的方法构建大鼠脾气虚证动物模型,分别进行D-木糖吸收实验及血常规检测。收集的尿液采用超高效液相色谱-飞行时间质谱联用(UPLC-Q-TOF-MS)分析,尿液代谢指纹图谱数据集进行主成分分析(PCA)和正交偏最小-判别分析(OPLSDA)等统计学分析,筛选脾气虚证的生物标记物。D-木糖和血常规测定结果表明脾气虚证大鼠造模成功。正、负离子模式下的PCA,OPLS-DA得分图显著区分模型组和空白组。根据OPLS-DA分析的S-plot图、VIP值、t检验及受试者特征曲线ROC下的面积(AUC),筛选并鉴定出苯丙氨酸、琥珀酸、乌头酸、异柠檬酸、甜菜碱、犬尿酸、吲哚、肌酸、肌酸酐、乳清酸、黄嘌呤、黄嘌呤酸等24个脾气虚证相关的生物标记物,主要涉及能量代谢、氨基酸代谢、色氨酸代谢、嘌呤代谢、嘧啶代谢。尿液代谢组学方法结合在线软件包进行数据处理解析代谢通路,可为脾气虚证及其他中医证候的研究提供新的方法和思路。
To identify biomarkers for spleen Qi deficiency by analyzing small molecule metabolites in urine,in order to expound the relationship between biomarkers and metabolic pathways. The spleen Qi deficiency model was established through dietary restriction and overstrain. All of the rats received D-xylose absorption experiment and blood routine test. Urine samples were collected in the next day.The urine samples were analyzed using UPLC-Q-TOF-MS to obtain the dataset of urine metabolic group. Principal component analysis(PCA),orthogonal partialleast squares-discriminant analysis(OPLS-DA) and other multivariate statistical methods were employed to evaluate the quality of the dataset and screen out potential biomarkers of spleen Qi deficiency. The results of D-xylose absorption and blood routine demonstrated that the spleen Qi deficiency model was successfully established. In positive ion mode and negative ion mode,PCA and OPLS-DA score plots could clearly distinguish model group and blank group. According to S-plot of OPLS-DA,VIP value,t-test and area under receiver operating characteristic curve( ROC),24 biomarkers,including phenylalanine,succinic acid,aconitic acid,isocitrate acid,betaine,kynurenine,indole,creatine,creatinine,orotic acid,xanthine,and xanthurenic acid,were identified as associated with the spleen Qi deficiency,mainly involving energy metabolism,amino acid metabolism,tryptophan metabolism,purine metabolism and pyrimidine metabolism. Urine metabolomics method combined with online software package for data processing and analysis metabolic pathway can provide new methods and ideas for studies for spleen Qi deficiency and other traditional Chinese medicine symptoms.

关键词(KeyWords): 脾气虚证;代谢组学;代谢指纹图谱;超高效液相色谱-飞行时间质谱联用;生物标记物
spleen Qi deficiency;metabolomics;metabolic fingerprint;UPLC-Q-TOF-MS;biomarker

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(81202917,81573607)

作者(Author): 刘武平;李婵艺;黄晶;廖婧竹;马文杰;陈宏远;芮雯;
LIU Wu-ping;LI Chan-yi;HUANG Jing;LIAO Jing-zhu;MA Wen-jie;CHEN Hong-yuan;RUI Wen;Central Laboratory,Guangdong Pharmaceutical University;Department of Immunology & Microbiology,Guangdong Pharmaceutical University;

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DOI: 10.19540/j.cnki.cjcmm.20170919.003

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