Biological Trace Element Research

, Volume 146, Issue 3, pp 402–409

Postnuclear Supernatant: An In Vitro Model for Assessing Cadmium-Induced Neurotoxicity

  • Namrata Govil
  • Shaista Chaudhary
  • Mohammad Waseem
  • Suhel Parvez


Cadmium (Cd) is a toxic heavy metal commonly found in industrial workplaces, a food contaminant and a major constituent of cigarette smoke. Most of the organs are susceptible to Cd-induced toxicity, including brain. Postnuclear supernatant (PNS) has been accepted as an in vitro model for assessing xenobiotic induced toxicity. The goal of the present study was to validate PNS as an in vitro model for investigating the effect of Cd-induced neurotoxicity. Neurotoxic induction by Cd was established in a dose-dependent manner in PNS in vitro. Enzymatic and non-enzymatic antioxidants were used as biomarkers of exposure. Antioxidant enzymatic activity was measured as a significant increase in activities of catalase, superoxide dismutase, and glutathione S-transferase. On exposure to Cd, a significant increase in acetylcholinesterase and decrease in sodium–potassium ATPase activity was also observed. Non-enzymatic effect was also demonstrated as a significant elevation in reduced glutathione and non-protein thiol activity, but there was no significant increase or decrease in the concentrations of protein thiol. In accordance with the toxicity of Cd towards the studied brain structure, Cd-induced oxidative stress has been a focus of toxicological research as a possible mechanism of neurotoxicity. Our results suggest that PNS preparations can be used as a model for future investigation of xenobiotic-induced neurotoxicity under in vitro conditions.


Cadmium Postnuclear supernatant Neurotoxicity Oxidative stress Biomarker Enzymatic antioxidants 


  1. 1.
    Patra RC, Rautray AK, Swarup D (2011) Oxidative stress in lead and cadmium toxicity and its amelioration. Vet Med Int. doi:10.4061/2011/457327
  2. 2.
    Nzengue Y, Candéias SM, Sauvaigo S, Douki T, Favier A, Rachidi W, Guiraud P (2011) The toxicity redox mechanisms of cadmium alone or together with copper and zinc homeostasis alteration: its redox biomarkers. J Trace Elem Med Biol 25:171–180PubMedCrossRefGoogle Scholar
  3. 3.
    Jiang G, Xu L, Zhang B, Wu L (2011) Effect of cadmium on proliferation and self-renewal activity of prostate stem/progenitor cells. Environ Toxicol Pharmacol 32:275–284PubMedCrossRefGoogle Scholar
  4. 4.
    Shimada H, Yasutake A, Hirashima T, Takamure Y, Kitano T, Waalkes MP, Imamura Y (2008) Strain difference of cadmium accumulation by liver slices of inbred Wistar-Imamichi and Fischer 344 rats. Toxicol In Vitro 22:338–343PubMedCrossRefGoogle Scholar
  5. 5.
    Flora SJ, Mittal M, Mehta A (2008) Heavy metal induced oxidative stress & its possible reversal by chelation therapy. Indian J Med Res 4:501–523Google Scholar
  6. 6.
    Méndez-Armenta M, Ríos C (2007) Cadmium neurotoxicity. Environ Toxicol Pharmacol 3:350–358CrossRefGoogle Scholar
  7. 7.
    Liu J, Qu W, Kadiiska MB (2009) Role of oxidative stress in cadmium toxicity and carcinogenesis. Toxicol Appl Pharmacol 3:209–214CrossRefGoogle Scholar
  8. 8.
    Joseph P (2009) Mechanisms of cadmium carcinogenesis. Toxicol Appl Pharmacol 3:272–279CrossRefGoogle Scholar
  9. 9.
    Hartwig A (2010) Mechanisms in cadmium-induced carcinogenicity: recent insights. Biometals 5:951–960CrossRefGoogle Scholar
  10. 10.
    Yazıhan N, Kaçar Koçak M, Akçıl E, Erdem O, Sayal A, Güven C, Akyürek N (2011) Involvement of galectin-3 in cadmium-induced cardiac toxicity. Anadolu Kardiyol Derg 11:479–484PubMedGoogle Scholar
  11. 11.
    López E, Arce C, Oset-Gasque MJ, Cañadas S, González MP (2006) Cadmium induces reactive oxygen species generation and lipid peroxidation in cortical neurons in culture. Free Radic Biol Med 40:940–951PubMedCrossRefGoogle Scholar
  12. 12.
    Rai A, Maurya SK, Khare P, Srivastava A, Bandyopadhyay S (2010) Characterization of developmental neurotoxicity of As, Cd, and Pb mixture: synergistic action of metal mixture in glial and neuronal functions. Toxicol Sci 2:586–601CrossRefGoogle Scholar
  13. 13.
    Fernandes CG, Borges CG, Seminotti B, Amaral AU, Knebel LA, Eichler P, de Oliveira AB, Leipnitz G, Wajner M (2011) Experimental evidence that methylmalonic acid provokes oxidative damage and compromises antioxidant defenses in nerve terminal and striatum of young rats. Cell Mol Neurobiol 5:775–785CrossRefGoogle Scholar
  14. 14.
    Calabrese V, Bates TE, Stella AMG (2000) NO synthase and NO-dependent signal pathways in brain aging and neurodegenerative disorders: the role of oxidant/antioxidant balance. Neurochem Res 25:1315–1341PubMedCrossRefGoogle Scholar
  15. 15.
    Milbury P, Blumberg JB (2003) Dietary antioxidants-human studies overview. In: Cutler RG, Rodriguez H (eds) Critical reviews of oxidative stress and aging: advances in basic science, diagnostics, and intervention. World Scientific, London, pp 487–502Google Scholar
  16. 16.
    Sgaravatti AM, Vargas BA, Zandoná BR, Deckmann KB, Rockenbach FJ, Moraes TB, Monserrat JM, Sgarbi MB, Pederzolli CD, Wyse AT, Wannmacher CM, Wajner M, Dutra-Filho CS (2008) Tyrosine promotes oxidative stress in cerebral cortex of young rats. Int J Dev Neurosci 26:551–559PubMedCrossRefGoogle Scholar
  17. 17.
    Moraes TB, Zanin F, da Rosa A, de Oliveira A, Coelho J, Petrillo F, Wajner M, Dutra-Filho CS (2010) Lipoic acid prevents oxidative stress in vitro and in vivo by an acute hyperphenylalaninemia chemically-induced in rat brain. J Neurol Sci 292:89–95PubMedCrossRefGoogle Scholar
  18. 18.
    Fernandes CG, Leipnitz G, Seminotti B, Amaral AU, Zanatta A, Vargas CR, Dutra Filho CS, Wajner M (2010) Experimental evidence that phenylalanine provokes oxidative stress in hippocampus and cerebral cortex of developing rats. Cell Mol Neurobiol 2:317–326CrossRefGoogle Scholar
  19. 19.
    Skrzycki M, Czeczot H, Majewska M, Podsiad M, Karlik W, Grono D, Wiechetek M (2010) Enzymatic antioxidant defense in isolated rat hepatocytes exposed to cadmium. Pol J Vet Sci 4:673–679Google Scholar
  20. 20.
    Clairborne A (1985) Catalase activity. In: Greenwald RA (ed) Handbook of methods for oxygen radical research. CRC, Boca Raton, pp 283–284Google Scholar
  21. 21.
    Misra HP, Fridovich I (1972) Role of superoxide anion in the autoxidation of epinephrine and a simple assay for superoxide dismutase. J Biol Chem 247:3170–3175PubMedGoogle Scholar
  22. 22.
    Habig WH, Pabst M, Jaoby WB (1974) Glutathione S-transferase: The first step in mercapturic acid formation. J Biol Chem 249:7130–7139PubMedGoogle Scholar
  23. 23.
    Ellman GL, Courtney KD, Andres V, Feather-Stone RM (1961) A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol 7:88–95PubMedCrossRefGoogle Scholar
  24. 24.
    Fiske CH, Subbarow YJ (1925) The calorimetric determination of phosphorus. Biol Chem 66:375–381Google Scholar
  25. 25.
    Jollow DJ, Mitchell JR, Zamppaglione Z, Gillette JR (1974) Bromobenzene induced liver necrosis; Protective role of glutathione and evidence for 3,4-bromobenzene oxide as the hepatotoxic metabolites. Pharmacol 11:151–169CrossRefGoogle Scholar
  26. 26.
    Sedlak J, Lindsay RH (1968) Estimation of total, protein-bound, and nonprotein sulfhydryl groups in tissue with Ellman’s reagent. Anal Biochem 25:192–205PubMedCrossRefGoogle Scholar
  27. 27.
    Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol. J Biol Chem 193:265–275PubMedGoogle Scholar
  28. 28.
    Wen YF, Zhao JQ, Bhadauria M, Nirala SK (2010) Pyridoxine mitigates cadmium induced hepatic cytotoxicity and oxidative stress. Environ Toxicol Pharmacol 30:169–174PubMedCrossRefGoogle Scholar
  29. 29.
    Templeton DM, Liu Y (2010) Multiple roles of cadmium in cell death and survival. Chem Biol Interact 188:267–275PubMedCrossRefGoogle Scholar
  30. 30.
    Urano S, Sato Y, Otonari T, Makabe S, Suzuki S, Ogata M, Endo T (1998) Aging and oxidative stress in neurodegeneration. Biofactors 7:103–112PubMedCrossRefGoogle Scholar
  31. 31.
    Slyshenkov VS, Shevalye AA, Liopo AV, Wojtczak L (2002) Protective role of l-methionine against free radical damage of rat brain synaptosomes. Acta Biochim Pol 4:907–916Google Scholar
  32. 32.
    Liu TY, Chen Y, Wang ZY, Ji LL, Wang ZT (2010) Pyrrolizidine alkaloid isoline-induced oxidative injury in various mouse tissues. Exp Toxicol Pathol 3:251–257CrossRefGoogle Scholar
  33. 33.
    Jurczuk M, Brzoska MM, Moniuszko-Jakoniuk J, Gałażyn-Sidorczuk M, Kulikowska-Karpińska E (2004) Antioxidant enzymes activity and lipid peroxidation in liver and kidney of rats exposed to cadmium and ethanol. Food Chem Toxicol 42:429–438PubMedCrossRefGoogle Scholar
  34. 34.
    Shagirtha K, Muthumani M, Prabu SM (2011) Melatonin abrogates cadmium induced oxidative stress related neurotoxicity in rats. Eur Rev Med Pharmacol Sci 15:1039–1050PubMedGoogle Scholar
  35. 35.
    Shukla PK, Khanna VK, Ali MM, Maurya RR, Handa SS, Srimal RC (2002) Protective effect of acorus calamus against acrylamide induced neurotoxicity. Phytother Res 16:256–260PubMedCrossRefGoogle Scholar
  36. 36.
    Carageorgiou H, Tzotzes V, Sideris A, Zarros A, Tsakiris S (2005) Cadmium effects on brain acetylcholinesterase activity and antioxidant status of adult rats: modulation by zinc, calcium and l-cysteine co-administration. Basic Clin Pharmacol Toxicol 97:320–324PubMedCrossRefGoogle Scholar
  37. 37.
    Mata M, Fink DJ, Gainer H, Smith CB, Davidsen L, Savakis H, Schwartz WJ, Sokoloff L (1980) Activity-dependent energy metabolism in rat posterior pituitary primarily reflects sodium pump activity. J Neurochem 34:213–215PubMedCrossRefGoogle Scholar
  38. 38.
    Hernandez R (1989) Brain Na+,K+-ATPase activity possibly regulated by a specific serotonin receptor. Brain Res 408:399–402CrossRefGoogle Scholar
  39. 39.
    Antonio MT, Corredor L, Leret ML (2003) Study of the activity of several brain enzymes like markers of the neurotoxicity induced by perinatal exposure to lead and/or cadmium. Toxicol Lett 143:331–340PubMedCrossRefGoogle Scholar
  40. 40.
    Carageorgiou H, Tzotzes V, Pantos C, Mourouzis C, Zarros A, Tsakiris S (2004) In vivo and in vitro effects of cadmium on adult rat brain total antioxidant status, acetylcholinesterase Na+,K+-ATPase and Mg2+-ATPase activities: protection by L-cysteine. Basic Clin Pharmacol Toxicol 94:112–118PubMedCrossRefGoogle Scholar
  41. 41.
    Zhu Y, Carvey PM, Ling Z (2006) Age-related changes in glutathione and glutathione-related enzymes in rat brain. Brain Res 1090:35–44PubMedCrossRefGoogle Scholar
  42. 42.
    Tabassum H, Parvez S, Rehman H, Dev Banerjee B, Siemen D, Raisuddin S (2007) Nephrotoxicity and its prevention by taurine in tamoxifen induced oxidative stress in mice. Hum Exp Toxicol 26:509–518PubMedCrossRefGoogle Scholar
  43. 43.
    Beyersmann D, Hechtenberg S (1997) Cadmium, gene regulation, and cellular signalling in mammalian cells. Toxicol Appl Pharmacol 144:247–261PubMedCrossRefGoogle Scholar
  44. 44.
    Schulz JB, Lindenau J, Seyfried J, Dichgans J (2000) Glutathione oxidative stress and neurodegeneration. Eur J Biochem 267:4904–4911PubMedCrossRefGoogle Scholar
  45. 45.
    Karihtala P, Soini Y (2007) Reactive oxygen species and antioxidant mechanisms in human tissues and their relation to malignancies. APMIS 115:81–103PubMedCrossRefGoogle Scholar
  46. 46.
    Hart RP, Rose CS, Hamer RM (1989) Neuropsychological effects of occupational exposure to cadmium. J Clin Exp Neuropsychol 11:933–943PubMedCrossRefGoogle Scholar
  47. 47.
    Webster WS, Valois AA (1981) The toxic effect of cadmium on the neonatal mouse CNS. J Neuropathol Exp Neurol 40:247–257PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Namrata Govil
    • 1
  • Shaista Chaudhary
    • 1
  • Mohammad Waseem
    • 1
  • Suhel Parvez
    • 1
  1. 1.Department of Medical Elementology and ToxicologyJamia Hamdard (Hamdard University)New DelhiIndia

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