基于高效液相色谱-飞行时间质谱技术的高血压病肝阳上亢证尿液代谢组学研究

Urine Metabonomic Study on Hypertension Patients of Ascendant Hyperactivity of Gan Yang Syndrome by High Performance Liquid Chromatography Coupled with Time of Flight Mass Spectrometry

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中文关键词: 代谢组学高效液相色谱-飞行时间质谱技术模式识别高血压病肝阳上亢证

英文关键词:metabolomics high performance liquid chromatography coupled with time of flight mass spectrometry pattern recognition hypertension of ascendant hyperactivity of Gan yang syndrome

基金项目:国家自然科学基金资助项目(No.30772865);教育部新世纪优秀人才支持计划资助项目(No.NCET-07-0522);山东省优秀中青年科学家奖励基金资助项目(No.2008BS03029);山东省2009—2010中医药科技发展计划资助项目(No.2009-018);山东省“泰山学者”建设工程专项经费资助项目

作者	单位
蒋海强	山东中医药大学实验中心
李运伦	山东中医药大学附属医院高血压中医临床研究基地
解君	山东中医药大学实验中心

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中文摘要:

目的研究高血压病肝阳上亢证患者的尿液代谢物变化,寻找其特征代谢物,探索高血压病肝阳上亢证的证候本质。方法选择高血压病肝阳上亢证患者10例为患者组,另设12名健康体检者为正常组,采用高效液相色谱-飞行时间质谱技术(HPLC-TOFMS)采集两组尿液代谢物谱,利用SIMCA-P软件进行主成分分析(principal component analysis,PCA)及偏最小二乘判别分析(partial least-square discriminant analysis,PLS-DA),寻找并鉴定尿液内差异代谢物,做出相关代谢途径的可能解释。结果 PCA方法处理患者组和正常组的尿液代谢物谱数据显示,两组HPLC-TOFMS谱数据可以明显分开。与正常组比较,患者组机体相关代谢发生显著变化。经PLS-DA确定与高血压病肝阳上亢证有关的代谢物,通过查询KEGG数据库共确定了15个化合物的结构及代谢途径,主要包括氨基酸、游离脂肪酸、神经鞘氨醇等。结论采用HPLC-TOFMS结合模式识别的代谢组学方法对高血压病肝阳上亢证患者尿液代谢物进行研究,从微观领域寻找小分子代谢标记物,有利于深入发现中医证候的生物学本质。

英文摘要:

Objective To study the changes of urine metabolites in hypertension patients of ascendant hyperactivity of Gan yang syndrome(AHGYS) ,and to explore its essence in hypertension patients.Methods Ten typical hypertension patients of AHGYS were recruited as the patient group,and the other twelve healthy volunteers were recruited as the normal group.The metabolite profiling in the urine were collected using by high performance liquid chromatography coupled with time of flight mass spectrometry(HPLC-TOFMS) .The principal component analysis(PCA) and partial least-square discriminant analysis(PLS-DA) were analyzed using SIMCA-P Software.The differential metabolites in the urine were found out and identified.The possible relevant metabolic pathways were explained.Results The data from the analysis by PCA in the urine samples of the patient group and the normal group showed,two sets of data could be obviously classified in the score plot.Compared with the normal group,significant changes happened to the body metabolism in the patient group.The metabolites relevant to hypertension patients of AHGYS were determined using the PLS-DA.Fifteen compounds of the structure and metabolic pathways had been confirmed through inquiring KEGG Database,mainly including amino acids,free fatty acids,sphingosine,and so on.Conclusions The hypertension patients of AHGYS were studied using HPLC-TOFMS combined with pattern recognition,thus finding out small molecular metabolic markers from the microscopic field,which was advantageous in probing the biological nature of Chinese medicine syndromes.

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