Ukr.Biochem.J. 2019; Volume 91, Issue 5, Sep-Oct, pp. 16-24


Agonists of CB1 and NMDA receptors decrease the toxic effect of organophosphorus compound paraoxon on PC12 cells

F. Salem1, F. Bahrami1,2, Z. Bahari2, Z. Jangravi3, S. Najafizadeh-Sari4

1Neuroscience Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran;
2Department of Physiology and Medical Physics, Faculty of Medicine, Baqiyatallah University of Medical Sciences, Tehran, Iran;
3Departmentof Biochemistry, Faculty of Medicine, Baqiyatallah University
of Medical Sciences, Tehran, Iran;
4Student’ Research Committee (SRC), Baqiyatallah University of Medical Sciences, Tehran, Iran;
e-mail: or

Received: 01 July 2019; Accepted: 13 August 2019

Pharmacological studies allow to suggest that activation of cannabinoid type 1 receptors (CB1) have a neuroprotective role against toxicity induced by organophosphate agents, but the exact mechanisms of this effect as well as interaction with receptors of other types are far from clear. Therefore, the aim of current study was to evaluate the effect of CB1 and NMDA receptors agonists on cell viability and biomarkers of oxidative stress and lipid peroxidation in PC12 cells exposed to paraoxon. PC12 cells were exposed to 100 µm paraoxon as organophosphate agent. Treatments with 1 µM arachidonyl-2′-chloroethylamide (ACEA) as specific agonist of CB1 receptors, 100 µM N-methyl-D-aspartate (NMDA) as agonist of NMDA receptors and 1 µM AM251 as antagonist of CB1 receptors were done. Cell via­bility and biomarkers of oxidative stress were evaluated after 48 h of incubation. The level of CB1 receptor protein was evaluated by Western blotting. It was demonstrated that PC12 cells treatment with paraoxon led to cell viability inhibition, glutathione level, superoxide dismutase and catalase activity reduction, lipid peroxidation intensification and CB1 receptor expression attenuation. Application of ACEA and NMDA was shown to be followed by normalization of these indices. The protective effect of ACEA was abolished when the CB1 receptors antagonist AM251 was applied. The study revealed that application of ACEA and NMDA can protect PC12 cells against paraoxon induced toxicity through antioxidant capacity increment, lipid peroxidation inhibition and enhanced expression of CB1 receptors.

Keywords: , , , , , ,


  1. Marsicano G, Lutz B. Neuromodulatory functions of the endocannabinoid system. J Endocrinol Invest. 2006;29(3 Suppl):27-46. PubMed
  2. Shohami E, Cohen-Yeshurun A, Magid L, Algali M, Mechoulam R. Endocannabinoids and traumatic brain injury. Br J Pharmacol. 2011 Aug;163(7):1402-10. PubMed, PubMedCentral, CrossRef
  3. Chen J, Lee CT, Errico S, Deng X, Cadet JL, Freed WJ. Protective effects of Delta(9)-tetrahydrocannabinol against N-methyl-d-aspartate-induced AF5 cell death. Brain Res Mol Brain Res. 2005 Apr 4;134(2):215-25. PubMed, PubMedCentral, CrossRef
  4. Coomber B, O’Donoghue MF, Mason R. Inhibition of endocannabinoid metabolism attenuates enhanced hippocampal neuronal activity induced by kainic acid. Synapse. 2008 Oct;62(10):746-55. PubMed, CrossRef
  5. Sinor AD, Irvin SM, Greenberg DA. Endocannabinoids protect cerebral cortical neurons from in vitro ischemia in rats. Neurosci Lett. 2000 Jan 14;278(3):157-60. PubMed, CrossRef
  6. Galve-Roperh I, Aguado T, Rueda D, Velasco G, Guzmán M. Endocannabinoids: a new family of lipid mediators involved in the regulation of neural cell development. Curr Pharm Des. 2006;12(18):2319-25. PubMed, CrossRef
  7. Basavarajappa BS, Nixon RA, Arancio O. Endocannabinoid system: emerging role from neurodevelopment to neurodegeneration. Mini Rev Med Chem. 2009 Apr;9(4):448-62. PubMed, PubMedCentral, CrossRef
  8. Bedse G, Romano A, Cianci S, Lavecchia AM, Lorenzo P, Elphick MR, Laferla FM, Vendemiale G, Grillo C, Altieri F, Cassano T, Gaetani S. Altered expression of the CB1 cannabinoid receptor in the triple transgenic mouse model of Alzheimer’s disease. J Alzheimers Dis. 2014;40(3):701-12. PubMed, CrossRef
  9. Campbell VA, Gowran A. Alzheimer’s disease; taking the edge off with cannabinoids? Br J Pharmacol. 2007 Nov;152(5):655-62. PubMed, PubMedCentral, CrossRef
  10. Van Laere K, Casteels C, Lunskens S, Goffin K, Grachev ID, Bormans G, Vandenberghe W. Regional changes in type 1 cannabinoid receptor availability in Parkinson’s disease in vivo. Neurobiol Aging. 2012 Mar;33(3):620.e1-8. PubMed, CrossRef
  11. Walsh S, Mnich K, Mackie K, Gorman AM, Finn DP, Dowd E. Loss of cannabinoid CB1 receptor expression in the 6-hydroxydopamine-induced nigrostriatal terminal lesion model of Parkinson’s disease in the rat. Brain Res Bull. 2010 Apr 5;81(6):543-8. PubMed, PubMedCentral, CrossRef
  12. Iuvone T, Esposito G, Esposito R, Santamaria R, Di Rosa M, Izzo AA. Neuroprotective effect of cannabidiol, a non-psychoactive component from Cannabis sativa, on beta-amyloid-induced toxicity in PC12 cells. J Neurochem. 2004 Apr;89(1):134-41. PubMed, CrossRef
  13. McAllister SD, Chan C, Taft RJ, Luu T, Abood ME, Moore DH, Aldape K, Yount G. Cannabinoids selectively inhibit proliferation and induce death of cultured human glioblastoma multiforme cells. J Neurooncol. 2005 Aug;74(1):31-40. PubMed, CrossRef
  14. Hillion JA, Takahashi K, Maric D, Ruetzler C, Barker JL, Hallenbeck JM. Development of an ischemic tolerance model in a PC12 cell line. J Cereb Blood Flow Metab. 2005 Feb;25(2):154-162. PubMed, PubMedCentral, CrossRef
  15. Vaudry D, Stork PJ, Lazarovici P, Eiden LE. Signaling pathways for PC12 cell differentiation: making the right connections. Science. 2002 May 31;296(5573):1648-9. PubMed, CrossRef
  16. Garzón J, de la Torre-Madrid E, Rodríguez-Muñoz M, Vicente-Sánchez A, Sánchez-Blázquez P. Gz mediates the long-lasting desensitization of brain CB1 receptors and is essential for cross-tolerance with morphine. Mol Pain. 2009 Mar 10;5:11. PubMed, PubMedCentral, CrossRef
  17. Sánchez-Blázquez P, Rodríguez-Muñoz M, Vicente-Sánchez A, Garzón J. Cannabinoid receptors couple to NMDA receptors to reduce the production of NO and the mobilization of zinc induced by glutamate. Antioxid Redox Signal. 2013 Nov 20;19(15):1766-82. PubMed, PubMedCentral, CrossRef
  18. Marchalant Y, Cerbai F, Brothers HM, Wenk GL. Cannabinoid receptor stimulation is anti-inflammatory and improves memory in old rats. Neurobiol Aging. 2008 Dec;29(12):1894-901. PubMed, PubMedCentral, CrossRef
  19. Salio C, Fischer J, Franzoni MF, Conrath M. Pre- and postsynaptic localizations of the CB1 cannabinoid receptor in the dorsal horn of the rat spinal cord. Neuroscience. 2002;110(4):755-64. PubMed, CrossRef
  20. Köfalvi A, Rodrigues RJ, Ledent C, Mackie K, Vizi ES, Cunha RA, Sperlágh B. Involvement of cannabinoid receptors in the regulation of neurotransmitter release in the rodent striatum: a combined immunochemical and pharmacological analysis. J Neurosci. 2005 Mar 16;25(11):2874-84. PubMed, CrossRef
  21. Rodriguez JJ, Mackie K, Pickel VM. Ultrastructural localization of the CB1 cannabinoid receptor in mu-opioid receptor patches of the rat Caudate putamen nucleus. J Neurosci. 2001 Feb 1;21(3):823-33. PubMed, CrossRef
  22. Chakraborty A, Murphy S, Coleman N. The Role of NMDA Receptors in Neural Stem Cell Proliferation and Differentiation. Stem Cells Dev. 2017 Jun 1;26(11):798-807.  PubMed, CrossRef
  23. Nacher J, McEwen BS. The role of N-methyl-D-asparate receptors in neurogenesis. Hippocampus. 2006;16(3):267-70. PubMed, CrossRef
  24. Wu X, Tian F, Okagaki P, Marini AM. Inhibition of N-methyl-D-aspartate receptors increases paraoxon-induced apoptosis in cultured neurons. Toxicol Appl Pharmacol. 2005 Oct 1;208(1):57-67. PubMed, PubMed
  25. Forrest D, Yuzaki M, Soares HD, Ng L, Luk DC, Sheng M, Stewart CL, Morgan JI, Connor JA, Curran T. Targeted disruption of NMDA receptor 1 gene abolishes NMDA response and results in neonatal death. Neuron. 1994 Aug;13(2):325-38. PubMed, CrossRef
  26. Tashiro A, Sandler VM, Toni N, Zhao C, Gage FH. NMDA-receptor-mediated, cell-specific integration of new neurons in adult dentate gyrus. Nature. 2006 Aug 24;442(7105):929-33. PubMed, CrossRef
  27. Girón-Pérez MI, Santerre A, Gonzalez-Jaime F, Casas-Solis J, Hernández-Coronado M, Peregrina-Sandoval J, Takemura A, Zaitseva G. Immunotoxicity and hepatic function evaluation in Nile tilapia (Oreochromis niloticus) exposed to diazinon. Fish Shellfish Immunol. 2007 Oct;23(4):760-9. PubMed, CrossRef
  28. Abdou HM, ElMazoudy RH. Oxidative damage, hyperlipidemia and histological alterations of cardiac and skeletal muscles induced by different doses of diazinon in female rats. J Hazard Mater. 2010 Oct 15;182(1-3):273-8. PubMed, CrossRef
  29. Kaur R, Sandhu HS. In vivo changes in antioxidant system and protective role of selenium in chlorpyrifos-induced subchronic toxicity in bubalus bubalis. Environ Toxicol Pharmacol. 2008 Jul;26(1):45-8.  PubMed, CrossRef
  30. Hashemi M, Bahrami F, Sahraei H, Golmanesh L, Sadri S. The neuroprotective effect of cannabinoid receptor agonist (WIN55,212-2) in paraoxon induced neurotoxicity in PC12 cells and N-methyl-D-aspartate receptor interaction. Yakhteh Med J. 2010;12(2):183-90.
  31. Winterbourn CC, Hawkins RE, Brian M, Carrell RW. The estimation of red cell superoxide dismutase activity. J Lab Clin Med. 1975 Feb;85(2):337-41. PubMed
  32. Aebi H. Catalase in vitro. Methods Enzymol. 1984;105:121-6. PubMed
  33. Tietze F. Enzymic method for quantitative determination of nanogram amounts of total and oxidized glutathione: applications to mammalian blood and other tissues. Anal Biochem. 1969 Mar;27(3):502-22. PubMed, CrossRef
  34. Satoh K. Serum lipid peroxide in cerebrovascular disorders determined by a new colorimetric method. Clin Chim Acta. 1978 Nov 15;90(1):37-43. PubMed, CrossRef
  35. Wolf SA, Bick-Sander A, Fabel K, Leal-Galicia P, Tauber S, Ramirez-Rodriguez G, Müller A, Melnik A, Waltinger TP, Ullrich O, Kempermann G. Cannabinoid receptor CB1 mediates baseline and activity-induced survival of new neurons in adult hippocampal neurogenesis. Cell Commun Signal. 2010 Jun 17;8:12. PubMed, PubMedCentral, CrossRef
  36. Ikonomidou C, Bosch F, Miksa M, Bittigau P, Vöckler J, Dikranian K, Tenkova TI, Stefovska V, Turski L, Olney JW. Blockade of NMDA receptors and apoptotic neurodegeneration in the developing brain. Science. 1999 Jan 1;283(5398):70-4. PubMed, CrossRef
  37. Lipton SA. NMDA receptor activity regulates transcription of antioxidant pathways. Nat Neurosci. 2008 Apr;11(4):381-2. PubMed, CrossRef
  38. Liu Z, Chen X, Gao Y, Sun S, Yang L, Yang Q, Bai F, Xiong L, Wang Q. Involvement of GluR2 up-regulation in neuroprotection by electroacupuncture pretreatment via cannabinoid CB1 receptor in mice. Sci Rep. 2015 Apr 1;5:9490. PubMed, PubMedCentral, CrossRef
  39. Pazos MR, Mohammed N, Lafuente H, Santos M, Martínez-Pinilla E, Moreno E, Valdizan E, Romero J, Pazos A, Franco R, Hillard CJ, Alvarez FJ, Martínez-Orgado J. Mechanisms of cannabidiol neuroprotection in hypoxic-ischemic newborn pigs: role of 5HT(1A) and CB2 receptors. Neuropharmacology. 2013 Aug;71:282-91. PubMed, CrossRef
  40. Rajan TS, Giacoppo S, Iori R, De Nicola GR, Grassi G, Pollastro F, Bramanti P, Mazzon E. Anti-inflammatory and antioxidant effects of a combination of cannabidiol and moringin in LPS-stimulated macrophages. Fitoterapia. 2016 Jul;112:104-15. PubMed, CrossRef
  41. Prins JM, Chao CK, Jacobson SM, Thompson CM, George KM. Oxidative stress resulting from exposure of a human salivary gland cells to paraoxon: an in vitro model for organophosphate oral exposure. Toxicol In Vitro. 2014 Aug;28(5):715-21. PubMed, PubMedCentral, CrossRef
  42. Contino M, Capparelli E, Colabufo NA, Bush AI. Editorial: The CB2 Cannabinoid System: A New Strategy in Neurodegenerative Disorder and Neuroinflammation. Front Neurosci. 2017 Apr 6;11:196. PubMed, PubMedCentral, CrossRef
  43. Sadri S, Bahrami F, Khazaei M, Hashemi M, Asgari A. Cannabinoid receptor agonist WIN-55,212-2 protects differentiated PC12 cells from organophosphorus- induced apoptosis. Int J Toxicol. 2010 Mar-Apr;29(2):201-8. PubMed, CrossRef
  44. Bahrami F, Hashemi M, Khalili F, Hashemi J, Asgari A. Stimulation of CB1 Cannabinoid and NMDA Receptors Increases Neuroprotective Effect against Diazinon-Induced Neurotoxicity. Neurophysiology. 2013;45(5-6):433-40. CrossRef

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