Analytical and Bioanalytical Chemistry

, Volume 406, Issue 5, pp 1339–1354 | Cite as

Metabolic profiling of urine and blood plasma in rat models of drug addiction on the basis of morphine, methamphetamine, and cocaine-induced conditioned place preference

  • Kei Zaitsu
  • Izuru Miyawaki
  • Kiyoko Bando
  • Hiroshi Horie
  • Noriaki Shima
  • Munehiro Katagi
  • Michiaki Tatsuno
  • Takeshi Bamba
  • Takako Sato
  • Akira Ishii
  • Hitoshi Tsuchihashi
  • Koichi Suzuki
  • Eiichiro Fukusaki
Research Paper


The metabolic profiles of urine and blood plasma in drug-addicted rat models based on morphine (MOR), methamphetamine (MA), and cocaine (COC)-induced conditioned place preference (CPP) were investigated. Rewarding effects induced by each drug were assessed by use of the CPP model. A mass spectrometry (MS)-based metabolomics approach was applied to urine and plasma of MOR, MA, and COC-addicted rats. In total, 57 metabolites in plasma and 70 metabolites in urine were identified by gas chromatography–MS. The metabolomics approach revealed that amounts of some metabolites, including tricarboxylic acid cycle intermediates, significantly changed in the urine of MOR-addicted rats. This result indicated that disruption of energy metabolism is deeply relevant to MOR addiction. In addition, 3-hydroxybutyric acid, l-tryptophan, cystine, and n-propylamine levels were significantly changed in the plasma of MOR-addicted rats. Lactose, spermidine, and stearic acid levels were significantly changed in the urine of MA-addicted rats. Threonine, cystine, and spermidine levels were significantly increased in the plasma of COC-addicted rats. In conclusion, differences in the metabolic profiles were suggestive of different biological states of MOR, MA, and COC addiction; these may be attributed to the different actions of the drugs on the brain reward circuitry and the resulting adaptation. In addition, the results showed possibility of predict the extent of MOR addiction by metabolic profiling. This is the first study to apply metabolomics to CPP models of drug addiction, and we demonstrated that metabolomics can be a multilateral approach to investigating the mechanism of drug addiction.


Metabolomics Drug addiction conditioned place preference Morphine Methamphetamine Cocaine 

Supplementary material

216_2013_7234_MOESM1_ESM.pdf (119 kb)
ESM 1 (PDF 118 kb)


  1. 1.
    Camí J, Farré M (2003) Mechanisms of disease drug addiction. N Engl J Med 4:975–986CrossRefGoogle Scholar
  2. 2.
    Goldstein RZ, Volkow ND (2002) Drug addiction and its underlying neurobiological basis: neuroimaging evidence for the involvement of the frontal cortex. Am J Psychiatry 159:1642–1652CrossRefGoogle Scholar
  3. 3.
    Falcón E, McClung CA (2009) A role for the circadian genes in drug addiction. Neuropharmacology 56:91–96CrossRefGoogle Scholar
  4. 4.
    Li CY, Mao X, Wei L (2008) Genes and (common) pathways underlying drug addiction. PLoS Comput Biol 4:28–34CrossRefGoogle Scholar
  5. 5.
    Chen XL, Lu G, Gong YX, Zhao LG, Chen J, Chi ZQ, Yang YM, Chen Z, Li Q, Liu JG (2007) Expression changes of hippocampal energy metabolism enzymes contribute to behavioral abnormalities during chronic morphine treatment. Cell Res 17:689–700CrossRefGoogle Scholar
  6. 6.
    Nestler EJ (2005) Is there a common molecular pathway for addiction? Nat Neurosci 8:1445–1449CrossRefGoogle Scholar
  7. 7.
    Hollander JA, Amelio AL, Kocerha J, Bali P, Lu Q, Willoughby D, Wahlestedt C, Conkright MD, Kenny PJ (2010) Striatal microRNA controls cocaine intake through CREB signaling. Nature 466:197–202CrossRefGoogle Scholar
  8. 8.
    Dreyer JL (2010) New insights into the roles of microRNA in drug addiction and neuroplasticity. Genome Med 2:92–98CrossRefGoogle Scholar
  9. 9.
    Dettmer K, Aronov PA, Hammock BD (2007) Mass spectrometry-based metabolomics. Mass Spectrom Rev 26:51–78CrossRefGoogle Scholar
  10. 10.
    Kaddurah-Daouk R, Krishnan KRR (2009) Metabolomics: a global biochemical approach to the study of central nervous system diseases. Neuropsychopharmacology 34:173–186CrossRefGoogle Scholar
  11. 11.
    Pasikanti KK, Ho PC, Chan ECY (2008) Gas chromatography/mass spectrometry in metabolic profiling of biological fluids. J Chromatogr B 871:202–211CrossRefGoogle Scholar
  12. 12.
    Patkar AA, Rozen S, Mannelli P, Matson W, Pae CU, Krishnan KR, Kaddurah-Daouk R (2009) Alterations in tryptophan and purine metabolism in cocaine addiction: a metabolomic study. Psychopharmacology 206:479–489CrossRefGoogle Scholar
  13. 13.
    Mannelli P, Patkar A, Rozen S, Matson W, Krishnan R, Kaddurah-Daouk R (2009) Opioid use affects antioxidant activity and purine metabolism: preliminary results. Hum Psychopharmacol 24:666–675CrossRefGoogle Scholar
  14. 14.
    Deng Y, Bu Q, Hu Z, Deng P, Yan G, Duan J, Hu C, Zhou J, Shao X, Zhao J, Li Y, Zhu R, Zhao Y, Cen X (2012) 1H-nuclear magnetic resonance-based metabonomic analysis of brain in rhesus monkeys with morphine treatment and withdrawal intervention. J Neurosci Res 90:2154–2162CrossRefGoogle Scholar
  15. 15.
    Wu M, Sahbaie P, Zheng M, Lobato R, Boison D, Clark JD, Peltz AG (2013) Opiate-induced changes in brain adenosine levels and narcotic drug responses. Neuroscience 228:235–242CrossRefGoogle Scholar
  16. 16.
    Hodson MP, Dear GJ, Roberts AD, Haylock CL, Ball RJ, Plumb RS, Stumpf CL, Griffin JL, Haselden JN (2007) A gender-specific discriminator in Sprague–Dawley rat urine: the deployment of a metabolic profiling strategy for biomarker discovery and identification. Anal Biochem 362:182–192CrossRefGoogle Scholar
  17. 17.
    Slupsky CM, Rankin KN, Wagner J, Fu H, Chang D, Weljie AM, Saude EJ, Lix B, Adamko DJ, Shah S, Greiner R, Sykes BD, Marrie TJ (2007) Investigation of the effects of gender, diurnal variation, and age in human urinary metabolomic profiles. Anal Chem 79:6995–7004CrossRefGoogle Scholar
  18. 18.
    Chen J, Liang J, Wang G, Han J, Cui C (2005) Repeated 2 Hz peripheral electrical stimulations suppress morphine-induced CPP and improve spatial memory ability in rats. Exp Neurol 194:550–556CrossRefGoogle Scholar
  19. 19.
    Suzuki T, Misawa M (1995) Sertindole antagonizes morphine-, cocaine-, and methamphetamine-induced place preference in the rat. Life Sci 57:1277–1284CrossRefGoogle Scholar
  20. 20.
    Åberg M, Wada D, Wall E, Izenwasser S (2007) Effect of MDMA (ecstacy) on activity and cocaine conditioned place preference in adult and adolescent rats. Neurotoxicol Teratol 29:37–46CrossRefGoogle Scholar
  21. 21.
    Bando K, Kawahara R, Kunimatsu T, Sakai J, Kimura J, Funabashi H et al (2010) Influences of biofluid sample collection and handling procedures on GC–MS-based metabolomics studies. J Biosci Bioeng 110:491–499CrossRefGoogle Scholar
  22. 22.
    Saude EJ, Sykes BD (2007) Urine stability for metabolomics studies: effects of preparation and storage. Metabolomics 3:19–27CrossRefGoogle Scholar
  23. 23.
    Trygg J AJ, Gullberg J, Johansson AI, Jonsson P, Antti H, Marklund SL, Moritz T (2005) Extraction and GC–MS analysis of the human blood plasma metabolome. Anal Chem 77:8086–8094CrossRefGoogle Scholar
  24. 24.
    Shima N, Miyawaki I, Bando K, Horie H, Zaitsu K, Katagi M, Bamba T, Tsuchihashi H, Fukusaki E (2011) Influences of methamphetamine-induced acute intoxication on urinary and plasma metabolic profiles in the rat. Toxicology 287:29–37CrossRefGoogle Scholar
  25. 25.
    Zhang Q, Wang G, Du Y, Zhu L, J A (2007) GC–MS analysis of the rat urine for metabolomics research. J Chromatogr B 854:20–25CrossRefGoogle Scholar
  26. 26.
    Tsugawa H, Bamba T, Shinohara M, Nishiumi S, Yoshida M, Fukusaki E (2011) Practical non-targeted gas chromatography/mass spectrometry-based metabolomics platform for metabolic phenotype analysis. J Biosci Bioeng 112:292–298CrossRefGoogle Scholar
  27. 27.
    Balster RL (1991) Drug abuse potential evaluation in animals. Br J Add 86:1549–1558CrossRefGoogle Scholar
  28. 28.
    Tzschentke TM (1998) Measuring reward with the conditioned place preference paradigm: a comprehensive review of drug effects, recent progress and new issues. Prog Neurobiol 56:613–672CrossRefGoogle Scholar
  29. 29.
    Messing RB, Flinchbaugh C, Waymire JC (1978) Changes in brain tryptophan and tyrosine following acute and chronic morphine administration. Neuropharmacology 17:391–396CrossRefGoogle Scholar
  30. 30.
    Larson AA, Takemori AE (1977) Effect of narcotics on the uptake of serotonin precursors by the rat brain. J Pharmacol Exp Ther 200:216–223Google Scholar
  31. 31.
    Yang L, Sun ZS, Zhu Y (2007) Proteomics analysis of rat prefrontal cortex in three phases of morphine-induced conditioned place preference. J Proteome Res 6:2239–2247CrossRefGoogle Scholar
  32. 32.
    Zhu H, Rockhold RW, Ho IK (1998) The role of glutamate in physical dependence on opioids. Jpn J Pharmacol 76:1–14CrossRefGoogle Scholar
  33. 33.
    Stryer L (1988) Biochemistry. Freeman, NYGoogle Scholar
  34. 34.
    Kim SJ, Lyoo IK, Hwang J, Sung YH, Lee HY, Lee DS, Jeong DU, Renshaw PF (2005) Frontal glucose hypometabolism in abstinent methamphetamine users. Neuropsychopharmacology 30:1383–1391Google Scholar
  35. 35.
    Farrar WL, Ferris DK, Harel-Bellan A (1989) The molecular basis of immune cytokine action. Crit Rev Ther Drug Carrier Syst 5:229–261Google Scholar
  36. 36.
    Wang X, Zhao T, Qiu Y, Su M, Jiang T, Zhou M, Zhao A, Jia W (2008) Metabonomics approach to understanding acute and chronic stress in rat models. J Proteome Res 8:2511–2518CrossRefGoogle Scholar
  37. 37.
    Goeders NE (2002) Stress and cocaine addiction. J Pharmacol Exp Ther 301:785–789CrossRefGoogle Scholar
  38. 38.
    Sarnyai Z, Shaham Y, Heinrichs SC (2001) The role of corticotropin-releasing factor in drug addiction. Pharmacol Rev 53:209–243Google Scholar
  39. 39.
    Butte JC, Kakihara R, Farnham ML, Noble EP (1973) The relationship between brain and plasma corticosterone stress response in developing rats. Endocrinology 92:1775–9CrossRefGoogle Scholar
  40. 40.
    Pitman DL, Ottenweller JE, Natelson BH (1988) Plasma corticosterone levels during repeated presentation of two intensities of restraint stress: chronic stress and habitation. Physiol Behav 43:47–55CrossRefGoogle Scholar
  41. 41.
    London ED, Cascella NG, Wong DF, Phillips RL, Dannals RF, Links JM, Herning R, Grayson R, Jaffe JH, Wagner HN (1990) Cocaine-induced reduction of glucose utilization in human brain. Arch Gen Psychiatry 47:567–574CrossRefGoogle Scholar
  42. 42.
    Volkow ND, Fowler JS, Wolf AP, Hitzemann R, Dewey S, Bendriem B, Alpert R, Hoff A (1991) Changes in brain glucose metabolism in cocaine dependence and withdrawal. Am J Psychiatry 148:621–626Google Scholar
  43. 43.
    Lindgren F, Hansen B, Karcher W (1996) Model Validation by permutation tests: applications to variable selection. J Chemom 10:521–532CrossRefGoogle Scholar
  44. 44.
    Barr AM, Panenka WJ, MacEwan GW, Thornton AE, Lang DJ, Honer WG, Lecomte T (2006) The need for speed: an update on methamphetamine addiction. J Psychiatry Neurosci 31:301–313Google Scholar
  45. 45.
    Brown JM, Hanson GR, Fleckenstein AE (2001) Regulation of the vesicular monoamine transporter-2: a novel mechanism for cocaine and other psychostimulants. J Pharmacol Exp Ther 296:762–767Google Scholar
  46. 46.
    Khoushbouei H, Wang H, Lechleiter JD, Javitch JA, Galli A (2003) Amphetamine-induced dopamine efflux. A voltage-sensitive and intracellular Na+-dependent mechanism. J Biol Chem 278:12070–12077CrossRefGoogle Scholar
  47. 47.
    Schmitz Y, Lee CJ, Schmauss C, Gonon F, Sulzer D (2001) Amphetamine distorts stimulation-dependent dopamine overflow: effects on D2 autoreceptors, transporters, and synaptic vesicle stores. J Neurosci 21:5916–5924Google Scholar
  48. 48.
    Saunders C, Ferrer JV, Shi L, Chen J, Merrill G, Lamb ME, Leeb-Lundberg LMF, Carvelli L, Javitch JA, Galli A (2000) Amphetamine-induced loss of human dopamine transporter activity: An internalization-dependent and cocaine-sensitive mechanism. PNAS 97:6850–6855CrossRefGoogle Scholar
  49. 49.
    Izenwasser S (2004) The role of the dopamine transporter in cocaine abuse. Neurotox Res 6:379–383CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Kei Zaitsu
    • 1
    • 2
  • Izuru Miyawaki
    • 3
  • Kiyoko Bando
    • 3
  • Hiroshi Horie
    • 3
  • Noriaki Shima
    • 2
  • Munehiro Katagi
    • 2
  • Michiaki Tatsuno
    • 2
  • Takeshi Bamba
    • 4
  • Takako Sato
    • 5
  • Akira Ishii
    • 1
  • Hitoshi Tsuchihashi
    • 5
  • Koichi Suzuki
    • 5
  • Eiichiro Fukusaki
    • 4
  1. 1.Department of Legal Medicine and BioethicsNagoya University Graduate School of MedicineNagoyaJapan
  2. 2.Forensic Science Laboratory, Osaka Prefectural Police HeadquartersOsakaJapan
  3. 3.Safety Research Laboratories, Dainippon Sumitomo Pharma Co., Ltd, OsakaJapan
  4. 4.Department of Biotechnology, Graduate School of EngineeringOsaka UniversityOsakaJapan
  5. 5.Department of Legal Medicine, Osaka Medical CollegeTakatsuki CityJapan

Personalised recommendations