Metabolomics: Towards Understanding Traditional Chinese Medicine

Abstract

Metabolomics represent a global understanding of metabolite complement of integrated living systems and dynamic responses to the changes of both endogenous and exogenous factors and has many potential applications and advantages for the research of complex systems. As a systemic approach, metabolomics adopts a “top-down” strategy to reflect the function of organisms from the end products of the metabolic network and to understand metabolic changes of a complete system caused by interventions in a holistic context. This property agrees with the holistic thinking of Traditional Chinese Medicine (TCM), a complex medical science, suggesting that metabolomics has the potential to impact our understanding of the theory behind the evidence-based Chinese medicine. Consequently, the development of robust metabolomic platforms will greatly facilitate, for example, the understanding of the action mechanisms of TCM formulae and the analysis of Chinese herbal (CHM) and mineral medicine, acupuncture, and Chinese medicine syndromes. This review summarizes some of the applications of metabolomics in special TCM issues with an emphasis on metabolic biomarker discovery.

metabolomics - biomarkers - traditional Chinese medicine - systems biology - NMR - molecular markers - LC‐MS - metabolic diseases

Reference

• 1 Oliver S G, Winson M K, Kell D B, Baganz F. Systematic functional analysis of the yeast genome.  Trends Biotechnol. 1998;  16 373-378
• 2 Workentine M L, Harrison J J, Weljie A M, Tran V A, Stenroos P U, Tremaroli V, Vogel H J, Ceri H, Turner R J. Phenotypic and metabolic profiling of colony morphology variants evolved from Pseudomonas fluorescens biofilms.  Environ Microbiol. 2010;  12 1565-1577
• 3 Pedersen K S, Kristensen T N, Loeschcke V, Petersen B O, Duus J Ø, Nielsen N C, Malmendal A. Metabolomic signatures of inbreeding at benign and stressful temperatures in Drosophila melanogaster.  Genetics. 2008;  180 1233-1243
• 4 Johansen K K, Wang L, Aasly J O, White L R, Matson W R, Henchcliffe C, Beal M F, Bogdanov M. Metabolomic profiling in LRRK2-related Parkinson's disease.  PLoS One. 2009;  4 e7551
• 5 Catchpole G S, Beckmann M, Enot D P, Mondhe M, Zywicki B, Taylor J, Hardy N, Smith A, King R D, Kell D B, Fiehn O, Draper J. Hierarchical metabolomics demonstrates substantial compositional similarity between genetically modified and conventional potato crops.  PNAS. 2005;  102 14458-14462
• 6 Nicholson J K, Lindon J C, Holmes E. ‘Metabonomics': understanding the metabolic responses of living systems to pathophysiological stimuli via multivariate statistical analysis of biological NMR spectroscopic data.  Xenobiotica. 1999;  29 1181-1189
• 7 Mally A, Amberg A, Hard G C, Dekant W. Are 4-hydroxy-2(E)-nonenal derived mercapturic acids and (1)H NMR metabonomics potential biomarkers of chemically induced oxidative stress in the kidney?.  Toxicology. 2007;  230 244-255
• 8 Stewart D, McDougall G J, Sungurtas J, Verrall S, Graham J, Martinussen I. Metabolomic approach to identifying bioactive compounds in berries: advances toward fruit nutritional enhancement.  Mol Nutr Food Res. 2007;  51 645-651
• 9 German J B, Roberts M A, Watkins S M. Genomics and metabolomics as markers for the interaction of diet and health: lessons from lipids.  J Nutr. 2003;  133 2078S-2083S
• 10 Schnackenberg L K, Beger R D. Monitoring the health to disease continuum with global metabolic profiling and systems biology.  Pharmacogenomics. 2006;  7 1077-1086
• 11 Pexa A, Boeger R H, Henle T, Schwedhelm E, Deussen A. Effects of moderate hyperhomocysteinaemia induced by 4 weeks methionine-enriched diet on metabolite profile and mesenteric artery function in rats.  Br J Nutr. 2008;  99 993-999
• 12 Astle J, Ferguson J T, German J B, Harrigan G G, Kelleher N L, Kodadek T, Parks B A, Roth M J, Singletary K W, Wenger C D, Mahady G B. Characterization of proteomic and metabolomic responses to dietary factors and supplements.  J Nutr. 2007;  137 2787-2793
• 13 Urakami K, Zangiacomi V, Yamaguchi K, Kusuhara M. Quantitative metabolome profiling of Illicium anisatum by capillary electrophoresis time-of-flight mass spectrometry.  Biomed Res. 2010;  31 161-163
• 14 Urayama S, Zou W, Brooks K, Tolstikov V. Comprehensive mass spectrometry based metabolic profiling of blood plasma reveals potent discriminatory classifiers of pancreatic cancer.  Rapid Commun Mass Spectrom. 2010;  24 613-620
• 15 Kim H K, Wilson E G, Choi Y H, Verpoorte R. Metabolomics: a tool for anticancer lead-finding from natural products.  Planta Med. 2010;  76 1094-1102
• 16 Sreekumar A, Poisson L M, Rajendiran T M, Khan A P, Cao Q, Yu J, Laxman B, Mehra R, Lonigro R J, Li Y, Nyati M K, Ahsan A, Kalyana-Sundaram S, Han B, Cao X, Byun J, Omenn G S, Ghosh D, Pennathur S, Alexander D C, Berger A, Shuster J R, Wei J T, Varambally S, Beecher C, Chinnaiyan A M. Metabolomic profiles delineate potential role for sarcosine in prostate cancer progression.  Nature. 2009;  457 910-914
• 17 Chan E C, Koh P K, Mal M, Cheah P Y, Eu K W, Backshall A, Cavill R, Nicholson J K, Keun H C. Metabolic profiling of human colorectal cancer using high-resolution magic angle spinning nuclear magnetic resonance (HR-MAS NMR) spectroscopy and gas chromatography mass spectrometry (GC/MS).  J Proteome Res. 2009;  8 352-361
• 18 Wang H, Ma C, Ma L, Du Z, Wang H, Ye H, Li G, Liu B, Xu G. Secondary metabolic profiling and artemisinin biosynthesis of two genotypes of Artemisia annua.  Planta Med. 2009;  75 1625-1633
• 19 Martins A M, Camacho D, Shuman J. A systems biology study of two distinct growth phases of Saccharomyces cerevisiae cultures.  Curr Genomics. 2004;  5 649-663
• 20 Lanza I R, Zhang S, Ward L E, Karakelides H, Raftery D, Nair K S. Quantitative metabolomics by H-NMR and LC-MS/MS confirms altered metabolic pathways in diabetes.  PLoS One. 2010;  5 e10538
• 21 Bao Y, Zhao T, Wang X, Qiu Y, Su M, Jia W. Metabonomic variations in the drug-treated type 2 diabetes mellitus patients and healthy volunteers.  J Proteome Res. 2009;  8 1623-1630
• 22 Lawaetz A J, Schmidt B, Staerk D, Jaroszewski J W, Bro R. Application of rotated PCA models to facilitate interpretation of metabolite profiles: commercial preparations of St. John's Wort.  Planta Med. 2009;  75 271-278
• 23 Goodacre R, Vaidyanathan S, Dunn W B, Harrigan G G, Kell D B. Metabolomics by numbers: acquiring and understanding global metabolite data.  Trends Biotechnol. 2004;  22 245-252
• 24 Want E J, Wilson I D, Gika H, Theodoridis G, Plumb R S, Shockcor J, Holmes E, Nicholson J K. Global metabolic profiling procedures for urine using UPLC-MS.  Nat Protoc. 2010;  5 1005-1018
• 25 Fiehn O. Metabolomics – the link between genotypes and phenotypes.  Plant Mol Biol. 2002;  48 155-171
• 26 Lao Y M, Jiang J G, Yan L. Application of metabonomic analytical techniques in the modernization and toxicology research of traditional Chinese medicine.  Br J Pharmacol. 2009;  157 1128-1141
• 27 Zhao J. Publishing Chinese medicine knowledge as linked data on the web.  Chin Med. 2010;  5 27
• 28 Beckonert O, Coen M, Keun H C, Wang Y, Ebbels T M, Holmes E, Lindon J C, Nicholson J K. High-resolution magic-angle-spinning NMR spectroscopy for metabolic profiling of intact tissues.  Nat Protoc. 2010;  5 1019-1032
• 29 Sobolev A P, Testone G, Santoro F, Nicolodi C, Iannelli M A, Amato M E, Ianniello A, Brosio E, Giannino D, Mannina L. Quality traits of conventional and transgenic lettuce (Lactuca sativa L.) at harvesting by NMR metabolic profiling.  J Agric Food Chem. 2010;  58 6928-6936
• 30 Straadt I K, Young J F, Bross P, Gregersen N, Oksbjerg N, Theil P K, Bertram H C. NMR-based metabonomic investigation of heat stress in myotubes reveals a time-dependent change in the metabolites.  J Agric Food Chem. 2010;  58 6376-6386
• 31 Straadt I K, Young J F, Petersen B O, Duus J Ø, Gregersen N, Bross P, Oksbjerg N, Bertram H C. Metabolic profiling of heat or anoxic stress in mouse C2C12 myotubes using multinuclear magnetic resonance spectroscopy.  Metabolism. 2010;  59 814-823
• 32 Kamleh M A, Dow J A, Watson D G. Applications of mass spectrometry in metabolomic studies of animal model and invertebrate systems.  Brief Funct Genomic Proteomic. 2009;  8 28-48
• 33 Moolenaar S H, Engelke U F, Wevers R A. Proton nuclear magnetic resonance spectroscopy of body fluids in the field of inborn errors of metabolism.  Ann Clin Biochem. 2003;  40 16-24
• 34 Field D, Sansone S A, Collis A, Booth T, Dukes P, Gregurick S K, Kennedy K, Kolar P, Kolker E, Maxon M, Millard S, Mugabushaka A M, Perrin N, Remacle J E, Remington K, Rocca-Serra P, Taylor C F, Thorley M, Tiwari B, Wilbanks J. Megascience. 'Omics data sharing.  Science. 2009;  326 234-236
• 35 Fiehn O. Extending the breadth of metabolite profiling by gas chromatography coupled to mass spectrometry.  Trends Anal Chem. 2008;  27 261-269
• 36 Tolstikov V V, Fiehn O, Tanaka N. Application of liquid chromatography-mass spectrometry analysis in metabolomics: reversed-phase monolithic capillary chromatography and hydrophilic chromatography coupled to electrospray ionization-mass spectrometry.  Methods Mol Biol. 2007;  358 141-155
• 37 Bollard M E, Stanley E G, Lindon J C, Nicholson J K, Holmes E. NMR-based metabonomic approaches for evaluating physiological influences on biofluid composition.  NMR Biomed. 2005;  18 143-162
• 38 Kell D B. Metabolomics and systems biology: making sense of the soup.  Curr Opin Microbiol. 2004;  7 296-307
• 39 Sajda P. Machine learning for detection and diagnosis of disease.  Annu Rev Biomed Eng. 2006;  8 537-565
• 40 Shin H, Markey M K. A machine learning perspective on the development of clinical decision support systems utilizing mass spectra of blood samples.  J Biomed Inform. 2006;  39 227-248
• 41 Patterson A D, Li H, Eichler G S, Krausz K W, Weinstein J N, Fornace Jr A J, Gonzalez F J, Idle J R. UPLC-ESI-TOFMS-based metabolomics and gene expression dynamics inspector self-organizing metabolomic maps as tools for understanding the cellular response to ionizing radiation.  Anal Chem. 2008;  80 665-674
• 42 Clayton T A, Lindon J C, Cloarec O, Antti H, Charuel C, Hanton G, Provost J P, Le Net J L, Baker D, Walley R J, Everett J R, Nicholson J K. Pharmaco-metabonomic phenotyping and personalized drug treatment.  Nature. 2006;  440 1073-1077
• 43 Roessner U, Bowne J. What is metabolomics all about?.  Biotechniques. 2009;  46 363-365
• 44 Rosenson R S. New technologies personalize diagnostics and therapeutics.  Curr Atheroscler Rep. 2010;  12 184-186
• 45 McClay J L, Adkins D E, Isern N G, O'Connell T M, Wooten J B, Zedler B K, Dasika M S, Webb B T, Webb-Robertson B J, Pounds J G, Murrelle E L, Leppert M F, van den Oord E J. (1)H nuclear magnetic resonance metabolomics analysis identifies novel urinary biomarkers for lung function.  J Proteome Res. 2010;  9 3083-3090
• 46 Wang X, Lv H, Sun H, Liu L, Yang B, Sun W, Wang P, Zhou D, Zhao L, Dou S, Zhang G, Cao H. Metabolic urinary profiling of alcohol hepatotoxicity and intervention effects of Yin Chen Hao Tang in rats using ultra-performance liquid chromatography/electrospray ionization quadruple time-of-flight mass spectrometry.  J Pharm Biomed Anal. 2008;  48 1161-1168
• 47 Wang P, Sun H, Lv H, Sun W, Yuan Y, Han Y, Wang D, Zhang A, Wang X. Thyroxine and reserpine-induced changes in metabolic profiles of rat urine and the therapeutic effect of Liu Wei Di Huang Wan detected by UPLC-HDMS.  J Pharm Biomed Anal. 2010;  53 631-645
• 48 Xie B, Gong T, Gao R, Liu J, Zuo J, Wang X, Zhang Z. Development of rat urinary HPLC-UV profiling for metabonomic study on Liuwei Dihuang pills.  J Pharm Biomed Anal. 2009;  49 492-497
• 49 Zhao X, Zhang Y, Meng X, Yin P, Deng C, Chen J, Wang Z, Xu G. Effect of a traditional Chinese medicine preparation Xindi soft capsule on rat model of acute blood stasis: a urinary metabonomics study based on liquid chromatography-mass spectrometry.  J Chromatogr B Analyt Technol Biomed Life Sci. 2008;  873 151-158
• 50 Lv Y, Liu X, Yan S, Liang X, Yang Y, Dai W, Zhang W. Metabolomic study of myocardial ischemia and intervention effects of Compound Danshen Tablets in rats using ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry.  J Pharm Biomed Anal. 2010;  52 129-135
• 51 Wang M, Rang W, Zhang Q, Huo C, Ma Z, Wang Y, Yan X, Gao Y. NMR-spectroscopy-based metabonomic approach to analysis of Siwutang, a novel prescription, treated blood deficiency in mice.  Zhongguo Zhong Yao Za Zhi. 2010;  35 630-634
• 52 Vlietinck A, Pieters L, Apers S. Legal requirements for the quality of herbal substances and herbal preparations for the manufacturing of herbal medicinal products in the European union.  Planta Med. 2009;  75 683-688
• 53 Chen M, Ni Y, Duan H, Qiu Y, Guo C, Jiao Y, Shi H, Su M, Jia W. Mass spectrometry-based metabolic profiling of rat urine associated with general toxicity induced by the multiglycoside of Tripterygium wilfordii Hook. f.  Chem Res Toxicol. 2008;  21 288-294
• 54 Liu Y, Huang R, Liu L, Peng J, Xiao B, Yang J, Miao Z, Huang H. Metabonomics study of urine from Sprague-Dawley rats exposed to Huang-yao-zi using (1)H NMR spectroscopy.  J Pharm Biomed Anal. 2010;  52 136-141
• 55 Xu G, Ma X, Zhang Q, Li B, Huang L, Yu R, Liu H. Study of metabonomics on pharmacological action appraisal Rhizoma coptidis in rats.  Zhongguo Zhong Yao Za Zhi. 2009;  34 1845-1847
• 56 Gu Y, Zhang Y, Shi X, Li X, Hong J, Chen J, Gu W, Lu X, Xu G, Ning G. Effect of traditional Chinese medicine berberine on type 2 diabetes based on comprehensive metabonomics.  Talanta. 2010;  81 766-772
• 57 Dai H, Xiao C, Liu H, Hao F, Tang H. Combined NMR and LC-DAD-MS analysis reveals comprehensive metabonomic variations for three phenotypic cultivars of Salvia miltiorrhiza Bunge.  J Proteome Res. 2010;  9 1565-1578
• 58 Agnolet S, Jaroszewski J W, Verpoorte R, Staerk D. H NMR-based metabolomics combined with HPLC-PDA-MS- SPE-NMR for investigation of standardized Ginkgo biloba preparations.  Metabolomics. 2010;  6 292-302
• 59 Zhang Q, Wang G J, A J Y, Wu D, Zhu L L, Ma B, Du Y. Application of GC/MS-based metabonomic profiling in studying the lipid-regulating effects of Ginkgo biloba extract on diet-induced hyperlipidemia in rats.  Acta Pharmacol Sin. 2009;  30 1674-1687
• 60 Liang Q, Ni C, Xie M, Zhang Q, Zhang Y X, Yan X Z, Yang M J, Peng S Q, Zhang Y Z. Nephrotoxicity study of Aristolochia fangchi in rats by metabonomics.  Zhong Xi Yi Jie He Xue Bao. 2009;  7 746-752
• 61 Chen M, Su M, Zhao L, Jiang J, Liu P, Cheng J, Lai Y, Liu Y, Jia W. Metabonomic study of aristolochic acid-induced nephrotoxicity in rats.  J Proteome Res. 2006;  5 995-1002
• 62 Jacob S S, Smith N W, Legido-Quigley C. Assessment of Chinese medicinal herb metabolite profiles by UPLC-MS- based methodology for the detection of aristolochic acids.  J Sep Sci. 2007;  30 1200-1206
• 63 Yan S, Wu B, Lin Z, Jin H, Huang J, Yang Y, Zhang X, Shen Z, Zhang W. Metabonomic characterization of aging and investigation on the anti-aging effects of total flavones of Epimedium.  Mol Biosyst. 2009;  5 1204-1213
• 64 Huang J H, Shen Z Y, Wu B. Effect and mechanism of Epimedium flavanoids for aging retardation from viewpoint of transcriptomics and metabonomics.  Zhongguo Zhong Xi Yi Jie He Za Zhi. 2008;  28 47-50
• 65 Wu B, Yan S, Lin Z, Wang Q, Yang Y, Yang G, Shen Z, Zhang W. Metabonomic study on ageing: NMR-based investigation into rat urinary metabolites and the effect of the total flavone of Epimedium.  Mol Biosyst. 2008;  4 855-861
• 66 Li F, Lu X, Liu H, Liu M, Xiong Z. A pharmaco-metabonomic study on the therapeutic basis and metabolic effects of Epimedium brevicornum Maxim. on hydrocortisone-induced rat using UPLC-MS.  Biomed Chromatogr. 2007;  21 397-405
• 67 Sun B, Li L, Wu S, Zhang Q, Li H, Chen H, Li F, Dong F, Yan X. Metabolomic analysis of biofluids from rats treated with Aconitum alkaloids using nuclear magnetic resonance and gas chromatography/time-of-flight mass spectrometry.  Anal Biochem. 2009;  395 125-133
• 68 Sun B, Wu S, Li L, Li H, Zhang Q, Chen H, Li F, Dong F, Yan X. A metabolomic analysis of the toxicity of Aconitum sp. alkaloids in rats using gas chromatography/mass spectrometry.  Rapid Commun Mass Spectrom. 2009;  23 1221-1228
• 69 Li L, Sun B, Zhang Q, Fang J, Ma K, Li Y, Chen H, Dong F, Gao Y, Li F, Yan X. Metabonomic study on the toxicity of Hei-Shun-Pian, the processed lateral root of Aconitum carmichaelii Debx. (Ranunculaceae).  J Ethnopharmacol. 2008;  116 561-568
• 70 Liu N Q, Cao M, Frédérich M, Choi Y H, Verpoorte R, van der Kooy F. Metabolomic investigation of the ethnopharmacological use of Artemisia afra with NMR spectroscopy and multivariate data analysis.  J Ethnopharmacol. 2010;  128 230-235
• 71 Wang H, Ma C, Ma L, Du Z, Wang H, Ye H, Li G, Liu B, Xu G. Secondary metabolic profiling and artemisinin biosynthesis of two genotypes of Artemisia annua.  Planta Med. 2009;  75 1625-1633
• 72 Xie G, Plumb R, Su M, Xu Z, Zhao A, Qiu M, Long X, Liu Z, Jia W. Ultra-performance LC/TOF MS analysis of medicinal Panax herbs for metabolomic research.  J Sep Sci. 2008;  31 1015-1026
• 73 Kang J, Lee S, Kang S, Kwon H N, Park J H, Kwon S W, Park S. NMR-based metabolomics approach for the differentiation of ginseng (Panax ginseng) roots from different origins.  Arch Pharm Res. 2008;  31 330-336
• 74 Wang Y, Wang J, Yao M, Zhao X, Fritsche J, Schmitt-Kopplin P, Cai Z, Wan D, Lu X, Yang S, Gu J, Häring H U, Schleicher E D, Lehmann R, Xu G. Metabonomics study on the effects of the ginsenoside Rg3 in a beta- cyclodextrin-based formulation on tumor-bearing rats by a fully automatic hydrophilic interaction/reversed-phase column-switching HPLC-ESI-MS approach.  Anal Chem. 2008;  80 4680-4688
• 75 Liang Y, Kang A, Xie T, Zheng X, Dai C, Hao H, A J, Sheng L, Xie L, Wang G J. Influence of segmental and selected ion monitoring on quantitation of multi-component using high-pressure liquid chromatography-quadrupole mass spectrometry: Simultaneous detection of 16 saponins in rat plasma as a case.  J Chromatogr A. 2010;  1217 4501-4506
• 76 Law W S, Huang P Y, Ong E S, Ong C N, Li S F, Pasikanti K K, Chan E C. Metabonomics investigation of human urine after ingestion of green tea with gas chromatography/mass spectrometry, liquid chromatography/mass spectrometry and (1)H NMR spectroscopy.  Rapid Commun Mass Spectrom. 2008;  22 2436-2446
• 77 Van Dorsten F A, Daykin C A, Mulder T P, Van Duynhoven J P. Metabonomics approach to determine metabolic differences between green tea and black tea consumption.  J Agric Food Chem. 2006;  54 6929-6938
• 78 Yi L Z, Yuan D L, Liang Y Z, Xie P S, Zhao Y. Fingerprinting alterations of secondary metabolites of tangerine peels during growth by HPLC-DAD and chemometric methods.  Anal Chim Acta. 2009;  649 43-51
• 79 Giri S, Idle J R, Chen C, Zabriskie T M, Krausz K W, Gonzalez F J. A metabolomic approach to the metabolism of the areca nut alkaloids arecoline and arecaidine in the mouse.  Chem Res Toxicol. 2006;  19 818-827
• 80 Adams M, Mahringer A, Kunert O, Fricker G, Efferth T, Bauer R. Cytotoxicity and p-glycoprotein modulating effects of quinolones and indoloquinazolines from the Chinese herb Evodia rutaecarpa.  Planta Med. 2007;  73 1554-1557
• 81 Zhang Q, Xu G, Wu L, Ma X, Zeng Z, Huang L, Yu R, Liu H. Preliminary study of metabonomics on aqueous extract of Evodia rutaecarpa in sprague-dawley rats.  Zhongguo Zhong Yao Za Zhi. 2010;  35 99-102
• 82 Xiao C, Dai H, Liu H, Wang Y, Tang H. Revealing the metabonomic variation of rosemary extracts using 1H NMR spectroscopy and multivariate data analysis.  J Agric Food Chem. 2008;  56 10142-10153
• 83 Wei L, Liao P, Wu H, Li X, Pei F, Li W, Wu Y. Toxicological effects of cinnabar in rats by NMR-based metabolic profiling of urine and serum.  Toxicol Appl Pharmacol. 2008;  227 417-429
• 84 Wei L, Liao P, Wu H, Li X, Pei F, Li W, Wu Y. Metabolic profiling studies on the toxicological effects of realgar in rats by (1)H NMR spectroscopy.  Toxicol Appl Pharmacol. 2009;  234 314-325
• 85 Liang F, Koya D. Acupuncture: is it effective for treatment of insulin resistance?.  Diabetes Obes Metab. 2010;  12 555-569
• 86 Wu Q, Zhang Q, Sun B, Yan X, Tang Y, Qiao X, Chen Q, Yu S, Liang F. (1)H NMR-based metabonomic study on the metabolic changes in the plasma of patients with functional dyspepsia and the effect of acupuncture.  J Pharm Biomed Anal. 2010;  51 698-704
• 87 Qiu Y, Chen M, Su M, Xie G, Li X, Zhou M, Zhao A, Jiang J, Jia W. Metabolic profiling reveals therapeutic effects of Herba Cistanches in an animal model of hydrocortisone-induced “kidney-deficiency syndrome”.  Chin Med. 2008;  3 3
• 88 Yan B, A J, Hao H, Wang G, Zhu X, Zha W, Liu L, Guan E, Zhang Y, Gu S, Huang Q, Zheng Y. Metabonomic phenotype and identification of “heart blood stasis obstruction pattern” and “qi and yin deficiency pattern” of myocardial ischemia rat models.  Sci China C Life Sci. 2009;  52 1081-1090
• 89 Chen M, Zhao L, Jia W. Metabonomic study on the biochemical profiles of a hydrocortisone-induced animal model.  J Proteome Res. 2005;  4 2391-2396
• 90 Luo H G, Ding J, Yue G X, Chen J X. Metabonomic study of syndrome of liver qi stagnation and spleen deficiency in rats.  Zhong Xi Yi Jie He Xue Bao. 2007;  5 307-313