Amino Acids

, Volume 46, Issue 9, pp 2219–2229 | Cite as

Metabolomic analysis of amino acid and energy metabolism in rats supplemented with chlorogenic acid

  • Zheng Ruan
  • Yuhui Yang
  • Yan Zhou
  • Yanmei Wen
  • Sheng Ding
  • Gang Liu
  • Xin Wu
  • Peng LiaoEmail author
  • Zeyuan Deng
  • Houssein Assaad
  • Guoyao Wu
  • Yulong YinEmail author
Original Article


This study was conducted to investigate effects of chlorogenic acid (CGA) supplementation on serum and hepatic metabolomes in rats. Rats received daily intragastric administration of either CGA (60 mg/kg body weight) or distilled water (control) for 4 weeks. Growth performance, serum biochemical profiles, and hepatic morphology were measured. Additionally, serum and liver tissue extracts were analyzed for metabolomes by high-resolution 1H nuclear magnetic resonance-based metabolomics and multivariate statistics. CGA did not affect rat growth performance, serum biochemical profiles, or hepatic morphology. However, supplementation with CGA decreased serum concentrations of lactate, pyruvate, succinate, citrate, β-hydroxybutyrate and acetoacetate, while increasing serum concentrations of glycine and hepatic concentrations of glutathione. These results suggest that CGA supplementation results in perturbation of energy and amino acid metabolism in rats. We suggest that glycine and glutathione in serum may be useful biomarkers for biological properties of CGA on nitrogen metabolism in vivo.


Chlorogenic acid Amino acids Metabolism Nuclear magnetic resonance spectroscopy 



Chlorogenic acid


Principal components


Principal component analysis


Nuclear magnetic resonance



This research was financially supported by National Natural Science Foundation of China (Grant No. 31001014), the Research Foundation (SKLF-TS-201108 and SKLF-TS-200817) and the Open Project Program (SKLF-KF-201005 and SKLF-KF-201216) of State Key Laboratory of Food Science and Technology at Nanchang University, and Texas A&M AgriLife Research H82000. H. Assaad was supported by a postdoctoral training grant (R25T-CA090301) from the National Cancer Institute.

Conflict of interest

The authors declare that they have no conflict of interests.


  1. Algamdi N, Mullen W, Crozier A (2011) Tea prepared from Anastatica hirerochuntica seeds contains a diversity of antioxidant flavonoids, chlorogenic acids and phenolic compounds. Phytochemistry 72:248–254PubMedCrossRefGoogle Scholar
  2. Cai Y, Luo Q, Sun M et al (2004) Antioxidant activity and phenolic compounds of 112 traditional Chinese medicinal plants associated with anticancer. Life Sci 74:2157–2184PubMedCrossRefGoogle Scholar
  3. Cruz M, Maldonado-Bernal C, Mondragon-Gonzalez R et al (2008) Glycine treatment decreases proinflammatory cytokines and increases interferon-gamma in patients with type 2 diabetes. J Endocrinol Invest 31:694–699PubMedCrossRefGoogle Scholar
  4. Dai ZL, Zhang J, Wu G et al (2010) Utilization of amino acids by bacteria from the pig small intestine. Amino Acids 39:1201–1215PubMedCrossRefGoogle Scholar
  5. Dai ZL, Wu G, Zhu WY (2011) Amino acid metabolism in intestinal bacteria: links between gut ecology and host health. Front Biosci 16:1768–1786CrossRefGoogle Scholar
  6. Dai ZL, Li XL, Xi PB et al (2012a) Metabolism of select amino acids in bacteria from the pig small intestine. Amino Acids 42:1597–1608PubMedCrossRefGoogle Scholar
  7. Dai ZL, Li XL, Xi PB et al (2012b) Regulatory role for l-arginine in the utilization of amino acids by pig small-intestinal bacteria. Amino Acids 43:233–244PubMedCrossRefGoogle Scholar
  8. Dai ZL, Wu ZL, Yang Y et al (2013a) Nitric oxide and energy metabolism in mammals. BioFactors 39:383–391PubMedCrossRefGoogle Scholar
  9. Dai ZL, Li XL, Xi PB et al (2013b) l-Glutamine regulates amino acid utilization by intestinal bacteria. Amino Acids 45:501–512PubMedCrossRefGoogle Scholar
  10. Dai ZL, Wu ZL, Jia SC et al (2014) Analysis of amino acid composition in proteins of animal tissues and foods as pre-column o-phthaldialdehyde derivatives by HPLC with fluorescence detection. J Chromatogr B. doi: 10.1016/j.jchromb.2014.03.025 Google Scholar
  11. Díaz-Flores M, Cruz M, Duran-Reyes G et al (2013) Oral supplementation with glycine reduces oxidative stress in patients with metabolic syndrome, improving their systolic blood pressure. Can J Physiol Pharmacol 91:855–860PubMedCrossRefGoogle Scholar
  12. El Hafidi M, Pérez I, Zamora J et al (2004) Glycine intake decreases plasma free fatty acids, adipose cell size, and blood pressure in sucrose-fed rats. Am J Physiol Regulatory Integrative Comp Physiol 287:1387–1393CrossRefGoogle Scholar
  13. Foo HL, Loh TC, Lai PW et al (2003) Effects of adding Lactobacillus plantarum I-UL4 metabolites in drinking water of rats. Pakistan J Nutr 2:283–288CrossRefGoogle Scholar
  14. Fu WJ, Stromberg AJ, Viele K et al (2010) Statistics and bioinformatics in nutritional sciences: analysis of complex data in the era of systems biology. J Nutr Biochem 21:561–572PubMedCentralPubMedCrossRefGoogle Scholar
  15. Greenberg JA, Boozer CN, Geliebter A (2006) Coffee, diabetes and weight control. Am J Clin Nutr 84:682–693PubMedGoogle Scholar
  16. Hayes JD, McLellan LI (1999) Glutathione and glutathione-dependent enzymes represent a co-ordinately regulated defence against oxidative stress. Free Radical Res 31:273–300CrossRefGoogle Scholar
  17. He ML, Wang YZ, Xu ZR et al (2003) Effect of dietary rare earth elements on growth performance and blood parameters of rats. J Anim Physiol Anim Nutr 87:229–235CrossRefGoogle Scholar
  18. He QH, Ren PP, Kong XF et al (2011) Metabolomics and its role in amino acid nutrition research. Front Biosci 16:2451–2460CrossRefGoogle Scholar
  19. He QH, Ren PP, Kong XF et al (2012) Comparison of serum metabolite compositions between obese and lean growing pigs using an NMR-based metabonomic approach. J Nutr Biochem 23:133–139PubMedCrossRefGoogle Scholar
  20. He LQ, Yang HS, Li TJ et al (2013) Effects of dietary l-lysine intake on the intestinal mucosa and expression of CAT genes in weaned piglets. Amino Acids 45:383–391PubMedCrossRefGoogle Scholar
  21. Hou YQ, Wang L, Zhang W et al (2012) Protective effects of N-acetylcysteine on intestinal functions of piglets challenged with lipopolysaccharide. Amino Acids 43:1233–1242PubMedCrossRefGoogle Scholar
  22. Hou YQ, Wang L, Yi D et al (2013) N-acetylcysteine reduces inflammation in the small intestine by regulating redox, EGF and TLR4 signaling. Amino Acids 45:513–522PubMedCrossRefGoogle Scholar
  23. Huang Z, Chang C (2008) Advances of study on glucose and lipids metabolism of chlorogenic acid regulating. J Hyg Res 37:637–639Google Scholar
  24. Ji L, Jiang P, Lu B et al (2013) Chlorogenic acid, a dietary polyphenol, protects acetaminophen-induced liver injury and its mechanism. J Nutr Biochem 24:1911–1919PubMedCrossRefGoogle Scholar
  25. Jiang CY, Yang KM, Yang L et al (2013) A 1H NMR-based metabonomic investigation of time-related metabolic trajectories of the plasma, urine and liver extracts of hyperlipidemic hamsters. PLoS One 8:e66786PubMedCentralPubMedCrossRefGoogle Scholar
  26. Jobgen WS, Fried SK, Fu WJ et al (2006) Regulatory role for the arginine-nitric oxide pathway in metabolism of energy substrates. J Nutr Biochem 17:571–588PubMedCrossRefGoogle Scholar
  27. Kasai H, Fukada S, Yamaizumi Z et al (2000) Action of chlorogenic acid in vegetables and fruits as an inhibitor of 8-hydroxydeoxyguanosine formation in vitro and in a rat carcinogenesis model. Food Chem Toxicol 38:467–471PubMedCrossRefGoogle Scholar
  28. Kauppinen RA, Nicholls DG (1986) Synaptosomal bioenergetics. The role of glycolysis, pyruvate oxidation and responses to hypoglycaemia. Eur J Biochem 158:159–165PubMedCrossRefGoogle Scholar
  29. Kong XF, Wu G, Yin YL (2011) Roles of phytochemicals in amino acid nutrition. Front Biosci S3:372–384CrossRefGoogle Scholar
  30. Krebs HA (1970) Rate control of the tricarboxylic acid cycle. Adv Enzym Regul 8:335–353CrossRefGoogle Scholar
  31. Krebs HA, Lowenstein JM (1960) The tricarboxylic acid cycle. Metab Pathw 1:129–203Google Scholar
  32. Li P, Yin YL, Li DF et al (2007) Amino acids and immune function. Br J Nutr 98:237–252PubMedCrossRefGoogle Scholar
  33. Li SY, Chang CQ, Ma FY et al (2009) Modulating effects of chlorogenic acid on lipids and glucose metabolism and expression of hepatic peroxisome proliferator-activated receptor-α in golden hamsters fed on high fat diet. Biomed Environ Sci 22:122–129PubMedCrossRefGoogle Scholar
  34. Li XL, Rezaei R, Li L et al (2011) Composition of amino acids in feed ingredients for animal diets. Amino Acids 40:1159–1168PubMedCrossRefGoogle Scholar
  35. Liao PQ, Wei L, Zhang XY et al (2007) Metabolic profiling of serum from gadolinium chloride-treated rats by 1H NMR spectroscopy. Anal Biochem 364:112–121PubMedCrossRefGoogle Scholar
  36. Lindon JC, Holmes E, Nicholson JK (2001) Pattern recognition methods and applications in biomedical magnetic resonance. Prog NMR Spectrosc 39:1–40CrossRefGoogle Scholar
  37. Liu G, Wang Y, Wang Z et al (2011) Nuclear magnetic resonance (NMR)-based metabolomic studies on urine and serum biochemical profiles after chronic cysteamine supplementation in rats. J Agric Food Chem 59:5572–5578PubMedCrossRefGoogle Scholar
  38. Lou Z, Wang H, Zhu S (2011) Antibacterial activity and mechanism of action of chlorogenic acid. J Food Sci 76:M398–M403PubMedCrossRefGoogle Scholar
  39. Mahaffey KR, Capar SG, Gladen BC et al (1981) Concurrent exposure to lead, cadmium, and arsenic. Effects on toxicity and tissue metal concentrations in the rat. J Lab Clin Med 98:463–481PubMedGoogle Scholar
  40. Marques V, Farah A (2009) Chlorogenic acids and related compounds in medicinal plants and infusions. Food Chem 113:1370–1376CrossRefGoogle Scholar
  41. Matilla B, Mauriz JL, Culebras JM et al (2002) Glycine: a cell-protecting anti-oxidant nutrient. Nutrición Hospitalaria 17:2–9PubMedGoogle Scholar
  42. Mauriz JL, Matilla B, Culebras JM et al (2001) Dietary glycine inhibits activation of nuclear factor kappa B and prevents liver injury in hemorrhagic shock in the rat. Free Radic Biol Med 31:1236–1244PubMedCrossRefGoogle Scholar
  43. Meng S, Cao J, Feng Q et al (2013) Roles of chlorogenic acid on regulating glucose and lipids metabolism: a review. Evid Based Complement Alternat Med. doi: 10.1155/2013/801457 Google Scholar
  44. Mubarak A, Hodgson JM, Considine MJ et al (2013) Supplementation of a high-fat diet with chlorogenic acid is associated with insulin resistance and hepatic lipid accumulation in mice. J Agric Food Chem 61:4371–4378PubMedCrossRefGoogle Scholar
  45. Park JB (2013) Isolation and quantification of major chlorogenic acids in three major instant coffee brands and their potential effects on H2O2-induced mitochondrial membrane depolarization and apoptosis in PC-12 cells. Food Funct 4:1632–1638PubMedCrossRefGoogle Scholar
  46. Ren WK, Luo W, Wu MM et al (2013a) Dietary l-glutamine supplementation improves pregnancy outcome in mice infected with type-2 porcine circovirus. Amino Acids 45:479–488PubMedCrossRefGoogle Scholar
  47. Ren WK, Zou LX, Ruan Z et al (2013b) Dietary l-proline supplementation confers immuno-stimulatory effects on inactivated Pasteurella multocida vaccine immunized mice. Amino Acids 45:555–561PubMedCrossRefGoogle Scholar
  48. Rezaei R, Wang WW, Wu ZL et al (2013a) Biochemical and physiological bases for utilization of dietary amino acids by young pigs. J Anim Sci Biotech 4:7CrossRefGoogle Scholar
  49. Rezaei R, Knabe DA, Tekwe CD et al (2013b) Dietary supplementation with monosodium glutamate is safe and improves growth performance in postweaning pigs. Amino Acids 44:911–923PubMedCrossRefGoogle Scholar
  50. Ruan Z, Lv Y, Fu X et al (2013) Metabolomic analysis of amino acid metabolism in colitic rats supplemented with lactosucrose. Amino Acids 45:877–887PubMedCrossRefGoogle Scholar
  51. Ruiz-Ramírez A, Ortiz-Balderas E, Cardozo-Saldaña G et al (2014) Glycine restores glutathione and protects against oxidative stress in vascular tissue from sucrose-fed rats. Clin Sci (Lond) 126:19–29CrossRefGoogle Scholar
  52. Satterfield MC, Wu G (2011) Growth and development of brown adipose tissue: significance and nutritional regulation. Front Biosci 16:1589–1608CrossRefGoogle Scholar
  53. Satterfield MC, Dunlap KA, Keisler DH et al (2012) Arginine nutrition and fetal brown adipose tissue development in diet-induced obese sheep. Amino Acids 43:1593–1603CrossRefGoogle Scholar
  54. Satterfield MC, Dunlap KA, Keisler DH et al (2013) Arginine nutrition and fetal brown adipose tissue development in nutrient-restricted sheep. Amino Acids 45:489–499PubMedCrossRefGoogle Scholar
  55. Schnackenberg L, Beger RD, Dragan Y (2005) NMR-based metabonomic evaluation of livers from rats chronically treated with tamoxifen, mestranol, and phenobarbital. Metabolomics 1:87–94CrossRefGoogle Scholar
  56. Schulz JB, Lindenau J, Seyfried J et al (2000) Glutathione, oxidative stress and neurodegeneration. Eur J Biochem 267:4904–4911PubMedCrossRefGoogle Scholar
  57. Shi H, Dong L, Bai Y et al (2009) Chlorogenic acid against carbon tetrachloride-induced liver fibrosis in rats. Eur J Pharmacol 623:119–124PubMedCrossRefGoogle Scholar
  58. Shi H, Dong L, Jiang J et al (2012) Chlorogenic acid reduces liver inflammation and fibrosis through inhibition of toll-like receptor 4 signaling pathway. Toxicology 303:107–114PubMedCrossRefGoogle Scholar
  59. Shi H, Dong L, Dang X et al (2013) Effect of chlorogenic acid on LPS-induced proinflammatory signaling in hepatic stellate cells. Inflamm Res 62:581–587PubMedCrossRefGoogle Scholar
  60. Tan BE, Li XG, Wu G et al (2012) Dynamic changes in blood flow and oxygen consumption in the portal-drained viscera of growing pigs receiving acute administration of l-arginine. Amino Acids 43:2481–2489PubMedCrossRefGoogle Scholar
  61. Trouillas P, Calliste CA, Allais DP et al (2003) Antioxidant, anti-inflammatory and antiproliferative properties of sixteen water plant extracts used in the Limousin countryside as herbal teas. Food Chem 80:399–407CrossRefGoogle Scholar
  62. Wan CW, Wong CNY, Pin WK et al (2013) Chlorogenic acid exhibits cholesterol lowering and fatty liver attenuating properties by up-regulating the gene expression of PPAR-α in hypercholesterolemic rats induced with a high-cholesterol diet. Phytother Res 27:545–551PubMedCrossRefGoogle Scholar
  63. Wang J, Wu G, Zhou H et al (2009) Emerging technologies for amino acid nutrition research in the post-genome era. Amino Acids 37:177–186PubMedCrossRefGoogle Scholar
  64. Wang QJ, Hou YQ, Yi D et al (2013a) Protective effects of N-acetylcysteine on alleviating acetic acid-induced colitis in a porcine model. BMC Gastroenterol 13:133PubMedCentralPubMedCrossRefGoogle Scholar
  65. Wang WW, Wu ZL, Dai ZL et al (2013b) Glycine metabolism in animals and humans: implications for nutrition and health. Amino Acids 45:463–477PubMedCrossRefGoogle Scholar
  66. Wang W, Dai Z, Wu Z et al (2014) Glycine is a nutritionally essential amino acid for maximal growth of milk-fed young pigs. Amino Acids. doi: 10.1007/s00726-014-1758-3
  67. Wei JW, Carroll RJ, Harden KK et al (2012) Comparisons of treatment means when factors do not interact in two-factorial studies. Amino Acids 42:2031–2035PubMedCentralPubMedCrossRefGoogle Scholar
  68. Wojtczak AB (1968) Control of acetoacetate and β-hydroxybutyrate production in rat liver mitochondria. Biochem Biophys Res Commun 31:634–640PubMedCrossRefGoogle Scholar
  69. Wu G (2010) Functional amino acids in growth, reproduction and health. Adv Nutr 1:31–37PubMedCentralPubMedCrossRefGoogle Scholar
  70. Wu G (2013a) Amino acids: biochemistry and nutrition. CRC Press, Boca RatonCrossRefGoogle Scholar
  71. Wu G (2013b) Functional amino acids in nutrition and health. Amino Acids 45:407–411PubMedCrossRefGoogle Scholar
  72. Wu GY, Gunasekara A, Brunengraber H et al (1991) Effects of extracellular pH, CO2, and \( {\text{HCO}}_{3}^{ - } \) on ketogenesis in perfused rat liver. Am J Physiol 261:E221–E226Google Scholar
  73. Wu GY, Fang YZ, Yang S et al (2004) Glutathione metabolism and its implications for health. J Nutr 134:489–492PubMedGoogle Scholar
  74. Wu G, Bazer FW, Burghardt RC et al (2011) Proline and hydroxyproline metabolism: implications for animal and human nutrition. Amino Acids 40:1053–1063PubMedCentralPubMedCrossRefGoogle Scholar
  75. Wu ZL, Satterfield MC, Bazer FW et al (2012) Regulation of brown adipose tissue development and white fat reduction by l-arginine. Curr Opin Clin Nutr Metab Care 15:529–538PubMedCrossRefGoogle Scholar
  76. Wu X, Zhang YZ, Yin YL et al (2013a) Roles of heat-shock protein 70 in protecting against intestinal mucosal damage. Front Biosci 18:356–365CrossRefGoogle Scholar
  77. Wu G, Wu ZL, Dai ZL et al (2013b) Dietary requirements of “nutritionally nonessential amino acids” by animals and humans. Amino Acids 44:1107–1113PubMedCrossRefGoogle Scholar
  78. Wu G, Bazer FW, Dai ZL et al (2014) Amino acid nutrition in animals: protein synthesis and beyond. Annu Rev Anim Biosci 2:387–417CrossRefGoogle Scholar
  79. Xu Y, Chen J, Yu X et al (2010) Protective effects of chlorogenic acid on acute hepatotoxicity induced by lipopolysaccharide in mice. Inflammation 59:871–877Google Scholar
  80. Xu XD, Zhang LH, Shao B et al (2013) Safety evaluation of meso-zeaxanthin. Food Control 32:678–686CrossRefGoogle Scholar
  81. Yang Y, Li C, Nie X et al (2007) Metabonomic studies of human hepatocellular carcinoma using high-resolution magic-angle spinning 1H NMR spectroscopy in conjunction with multivariate data analysis. J Proteome Res 6:2605–2614PubMedCrossRefGoogle Scholar
  82. Yao K, Guan S, Li T et al (2011) Dietary l-arginine supplementation enhances intestinal development and expression of vascular endothelial growth factor in weanling piglets. Br J Nutr 105:703–709PubMedCrossRefGoogle Scholar
  83. Yun N, Kang JW, Lee SM (2012) Protective effects of chlorogenic acid against ischemia/reperfusion injury in rat liver: molecular evidence of its antioxidant and anti-inflammatory properties. J Nutr Biochem 23:1249–1255PubMedCrossRefGoogle Scholar
  84. Zhang LT, Chang CQ, Liu Y et al (2011) Effect of chlorogenic acid on disordered glucose and lipid metabolism in db/db mice and its mechanism. Acta Academiae Medicinae Sinicae 33:281–286PubMedGoogle Scholar
  85. Zhao Y, Wang J, Ballevre O et al (2012) Antihypertensive effects and mechanisms of chlorogenic acids. Hypertens Res 35:370–402PubMedCrossRefGoogle Scholar
  86. Zira A, Kostidis S, Theocharis S et al (2013) 1H NMR-based metabonomics approach in a rat model of acute liver injury and regeneration induced by CCl4 administration. Toxicology 303:115–124PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Wien 2014

Authors and Affiliations

  • Zheng Ruan
    • 1
  • Yuhui Yang
    • 1
  • Yan Zhou
    • 1
  • Yanmei Wen
    • 1
  • Sheng Ding
    • 2
  • Gang Liu
    • 3
  • Xin Wu
    • 3
  • Peng Liao
    • 3
    • 4
    Email author
  • Zeyuan Deng
    • 1
  • Houssein Assaad
    • 5
  • Guoyao Wu
    • 5
  • Yulong Yin
    • 1
    • 3
    Email author
  1. 1.State Key Laboratory of Food Science and Technology, College of Life Science and Food EngineeringNanchang UniversityNanchangChina
  2. 2.Institute of Nutrition and Food SafetyCenter for Disease Control and Prevention of Jiangxi ProvinceNanchangChina
  3. 3.Hunan Engineering and Research Center of Animal and Poultry Science, Institute of Subtropical AgricultureChinese Academy of SciencesChangshaChina
  4. 4.Hunan New Wellful Co., LTDChangshaChina
  5. 5.Department of Animal ScienceTexas A&M UniversityCollege StationUSA

Personalised recommendations