Selegiline acts as neuroprotective agent against methamphetamine-prompted mood and cognitive related behavior and neurotoxicity in rats: Involvement of CREB/BDNF and Akt/GSK3 signal pathways

Document Type: Original Article


1 Department of Pharmaceutical Chemistry, Faculty of Pharmaceutical Chemistry, Pharmaceutical Sciences Branch, Islamic Azad University (IUAPS), Tehran, Iran

2 Razi Drug Research Center, Iran University of Medical Sciences, Tehran, Iran

3 Department of Medicinal Chemistry, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran

4 Department of medicine, Qom branch, Islamic Azad University, Iran



Objective(s): Present study investigated the neuroprotective effects of selegiline and the molecular mechanisms involved in methamphetamine-induced neurotoxicity.
Materials and Methods: Male wistar rats were randomly divided into six groups (10 rats in each group). Group 1 and group 2 received normal saline and methamphetamine (10 mg/kg), respectively. Groups 3, 4, 5 and 6 were treated simultaneously with methamphetamine and selegiline. From day 22 to day 28, forced swim test, elevated plus maze, and open field test were conducted to assess mood (anxiety and depression) levels, and from day 17 to day 21, Morris Water Maze was conducted for cognition assessment. On day 29, hippocampus of the animals were isolated and evaluated by ELISA method for oxidative, antioxidant, and inflammatory factors and expression levels of active (total) and inactive (phosphorylated) forms of cyclic AMP response element binding protein (CREB), brain-derived neurotrophic factor (BDNF), Akt (Protein Kinase B) and glycogen synthase kinase 3 (GSK3) proteins.
Results: Selegiline reduced behavioral impacts caused by methamphetamine in all doses. Methamphetamine administration may improve malondialdehyde, tumor necrosis factor-alpha, interleukin-1 beta and GSK3 (both forms). Moreover, methamphetamine reduced the activity of superoxide dismutase, glutathione peroxidase, glutathione reductase, amount of BDNF, CREB and Akt (both forms).
Conclusion: Current research showed that selegiline can protect the brain from methamphetamine-prompted neurodegeneration, and this could be intervened by CREB -BDNF or Akt-GSK3 signaling pathways.


1. Anglin MD, Burke C, Perrochet B, Stamper E, Dawud-Noursi S. History of the methamphetamine problem. J Psychoactive Drugs 2000; 32:137-141.
2. Graves SM, Xie Z, Zampese E, Stout KA, Tai RA, Schwarzschild SE, et al. Methamphetamine-induced mitochondrial oxidant stress mediated by monoamine oxidase metabolism of dopamine. FASEB J 2017;983-987.
3. Winslow BT, Voorhees KI, Pehl KA. Methamphetamine abuse. Am Fam Physician 2007; 76-80.
4. Rusyniak DE. Neurologic manifestations of chronic methamphetamine abuse. Psychiatr Clin North Am 2013; 36:261-275.
5. Figueira FH, Leal CQ, de Moraes Reis E, Röpke J, Wagner C, da Rocha JBT, et al. Effects of diphenyl diselenide on behavioral and biochemical changes induced by amphetamine in mice. J Neural Transm (Vienna) 2015; 122:201-209.
6. Peleg-Raibstein D, Sydekum E, Russig H, Feldon J. Withdrawal from repeated amphetamine administration leads to disruption of prepulse inhibition but not to disruption of latent inhibition. J Neural Transm (Vienna) 2006; 113:1323-1336.
7. Morton WA, Stockton GG. Methylphenidate abuse and psychiatric side effects. Prim Care Companion J Clin Psychiatry 2000; 2:159-164.
8. Barrett SP, Pihl RO. Oral methylphenidate-alcohol co-abuse. J Clin Psychopharmacol 2002; 22:633-634.
9. Klein-Schwartz W. Abuse and toxicity of methylphenidate. Curr Opin Pediatr 2002; 14:219-223.
10. Cadet JL, Krasnova IN. Molecular bases of methamphetamine-induced neurodegeneration. Int Rev Neurobiol 2009; 88:101-119.
11. Krasnova IN, Cadet JL. Methamphetamine toxicity and messengers of death. Brain Res Rev 2009; 60:379-407.
12. Pålhagen S, Heinonen E, Hägglund J, Kaugesaar T, Mäki-Ikola O, Palm R. Selegiline slows the progression of the symptoms of Parkinson disease. Neurology 2006; 66:1200-1206.
13. Shimazu S, Minami A, Kusumoto H, Yoneda F. Antidepressant-like effects of selegiline in the forced swim test. Eur Neuropsychopharmacol 2005; 15:563-571.
14. Beata C, Beaumont-Graff E, Diaz C, Marion M, Massal N, Marlois N, et al. Effects of alpha-casozepine (Zylkene) versus selegiline hydrochloride (Selgian, Anipryl) on anxiety disorders in dogs. Journal of Veterinary Behavior: Clinical Applications and Research 2007; 2:175-183.
15. Behrend EN. Update on drugs used to treat endocrine diseases in small animals. Vet Clin North Am Small Anim Pract 2006; 36:1087-1105.
16. Youdim MB. The path from anti Parkinson drug selegiline and rasagiline to multifunctional neuroprotective anti Alzheimer drugs ladostigil and m30. Curr Alzheimer Res 2006; 3:541-550.
17. Takahata K, Shimazu S, Katsuki H, Yoneda F, Akaike A. Effects of selegiline on antioxidant systems in the nigrostriatum in rat. Neural Transm (Vienna) 2006; 113:151-158.
18. Parvizpour A, Charkhpour M, Habibi-asl B, Shakhsi M, Ghaderi M, Hassanzadeh K. Repeated central administration of selegiline attenuated morphine physical dependence in rat. Pharmacol Rep 2013; 65:593-599.
19. Sofuoglu M, Kosten TR. Novel approaches to the treatment of cocaine addiction. CNS drugs 2005; 19:13-25.
20. Robinson DS. Dopamine and Depression. Primary Psychiatry 2007; 14:21-23.
21. Newton TF, De La Garza R, Fong T, Chiang N, Holmes TH, Bloch DA, et al. A comprehensive assessment of the safety of intravenous methamphetamine administration during treatment with selegiline. Pharmacol Biochem Behav 2005; 82:704-711.
22. Blendy JA. The role of CREB in depression and antidepressant treatment. Biol Psychiatry 2006; 59:1144-1150.
23. Lee B, Butcher GQ, Hoyt KR, Impey S, Obrietan K. Activity-dependent neuroprotection and cAMP response element-binding protein (CREB): kinase coupling, stimulus intensity, and temporal regulation of CREB phosphorylation at serine 133. J Neurosci 2005; 25:1137-1148.
24. Motaghinejad M, Motevalian M, Falak R, Heidari M, Sharzad M, Kalantari E. Neuroprotective effects of various doses of topiramate against methylphenidate-induced oxidative stress and inflammation in isolated rat amygdala: the possible role of CREB/BDNF signaling pathway. Neural Transm (Vienna) 2016; 123:1463-1477.
25. Motaghinejad M, Motevalian M, Babalouei F, Abdollahi M, Heidari M, Madjd Z. Possible involvement of CREB/BDNF signaling pathway in neuroprotective effects of topiramate against methylphenidate induced apoptosis, oxidative stress and inflammation in isolated hippocampus of rats: molecular, biochemical and histological evidences. Brain Res Bull 2017; 132:82-98.
26.Carlezon WA, Duman RS, Nestler EJ. The many faces of CREB. Trends Neurosci 2005; 28:436-445.
27. Kitagawa K. CREB and cAMP response element‐mediated gene expression in the ischemic brain. FEBS J 2007; 274:3210-3217.
28. Réus GZ, Stringari RB, Ribeiro KF, Ferraro AK, Vitto MF, Cesconetto P, et al. Ketamine plus imipramine treatment induces antidepressant-like behavior and increases CREB and BDNF protein levels and PKA and PKC phosphorylation in rat brain. Behav Brain Res 2011; 221:166-171.
29. Aguiar AS, Castro AA, Moreira EL, Glaser V, Santos AR, Tasca CI, et al. Short bouts of mild-intensity physical exercise improve spatial learning and memory in aging rats: involvement of hippocampal plasticity via AKT, CREB and BDNF signaling. Mech Ageing Dev 2011; 132:560-567.
30. Terranova C, Rizzo V, Rajan T, Naro A, Ahmad A, Ghilardi M, et al. Repetitive transcranial magnetic stimulation induces neuroprotection via activating CaMKIIα-CREB-Bcl-2 pathway in rat brain. Clinical Neurophysiology 2016; 127:e146.
31. Motaghinejad M, Motevalian M, Fatima S, Faraji F, Mozaffari S. The Neuroprotective Effect of Curcumin Against Nicotine-Induced Neurotoxicity is Mediated by CREB–BDNF Signaling Pathway. Neurochem Res 2017; 42:2921-2932.
32. Motaghinejad M, Motevalian M, Fatima S, Hashemi H, Gholami M. Curcumin confers neuroprotection against alcohol-induced hippocampal neurodegeneration via CREB-BDNF pathway in rats. Biomed Pharmacother 2017; 87:721-740.
33. Chen PC, Lao CL, Chen JC. Dual alteration of limbic dopamine D1 receptor‐mediated signalling and the Akt/GSK3 pathway in dopamine D3 receptor mutants during the development of methamphetamine sensitization. J Neurochem 2007; 100:225-241.
34. Motaghinejad M, Seyedjavadein Z, Motevalian M, Asadi M. The neuroprotective effect of lithium against high dose methylphenidate: possible role of BDNF. Neurotoxicology 2016; 56:40-54.
35. Dave KR, Saul I, Busto R, Ginsberg MD, Sick TJ, Pérez-Pinzón MA. Ischemic preconditioning preserves mitochondrial function after global cerebral ischemia in rat hippocampus. J Cereb Blood Flow Metab 2001;21:1401-1410.
36. Gould TD, Dao DT, Kovacsics CE. The open field test.  Mood and anxiety related phenotypes in mice. Springer; 2009. P. 1-20.
37. File SE, Lippa AS, Beer B, Lippa MT. Animal tests of anxiety. Curr Protoc Neurosci 2004; 26: 1-8.
38. Slattery DA, Cryan JF. Using the rat forced swim test to assess antidepressant-like activity in rodents. Nat Protoc 2012; 7:1009-1021.
39. Castagné V, Moser P, Roux S, Porsolt RD. Rodent models of depression: forced swim and tail suspension behavioral despair tests in rats and mice. Curr Protoc Pharmacol 2010; 49:1-8.
40. Walf AA, Frye CA. The use of the elevated plus maze as an assay of anxiety-related behavior in rodents. Nat Protoc 2007; 2:322-329.
41. Morris RG. Morris water maze. Scholarpedia 2008; 3:6315-6319.
42. Motaghinejad M, Fatima S, Karimian M, Ganji S. Protective effects of forced exercise against nicotine-induced anxiety, depression and cognition impairment in rat. J Basic Clin Physiol Pharmacol 2016; 27:19-27.
43. Arias C, Montiel T, Quiroz-Báez R, Massieu L. β-Amyloid neurotoxicity is exacerbated during glycolysis inhibition and mitochondrial impairment in the rat hippocampus in vivo and in isolated nerve terminals: implications for Alzheimer’s disease. Exp Neurol 2002; 176:163-174.
44. Weydert CJ, Cullen JJ. Measurement of superoxide dismutase, catalase and glutathione peroxidase in cultured cells and tissue. Nat Protoc 2010; 5:51-55.
45. Motaghinejad M, Motevalian M, Shabab B, Fatima S. Effects of acute doses of methylphenidate on inflammation and oxidative stress in isolated hippocampus and cerebral cortex of adult rats. J Neural Transm (Vienna) 2017;124:121-131.
46. Dong F, Zhang X, Culver B, Chew HG, Kelley RO, Ren J. Dietary iron deficiency induces ventricular dilation, mitochondrial ultrastructural aberrations and cytochrome c release: involvement of nitric oxide synthase and protein tyrosine nitration. Clinical Sci 2005; 109:277-286.
47. Garcia YJ, Rodríguez-Malaver AJ, Peñaloza N. Lipid peroxidation measurement by thiobarbituric acid assay in rat cerebellar slices. J Neurosci Methods 2005; 144:127-135.
48.Mousavi SN, Faghihi A, Motaghinejad M, Shiasi M, Imanparast F, Amiri HL, et al. Zinc and selenium co-supplementation reduces some lipid peroxidation and angiogenesis markers in a rat model of NAFLD-fed high fat diet. Biol Trace Elem Res 2018; 181:288-295.
49. Parikh V, Khan MM, Mahadik SP. Differential effects of antipsychotics on expression of antioxidant enzymes and membrane lipid peroxidation in rat brain. J Psychiatr Res 2003; 37:43-51.
50. Yannarelli GG, Fernández-Alvarez AJ, Santa-Cruz DM, Tomaro ML. Glutathione reductase activity and isoforms in leaves and roots of wheat plants subjected to cadmium stress. Phytochemistry 2007; 68:505-512.
51. Box A, Sureda A, Galgani F, Pons A, Deudero S. Assessment of environmental pollution at Balearic Islands applying oxidative stress biomarkers in the mussel Mytilus galloprovincialis. Comp Biochem Physiol C Toxicol Pharmacol 2007; 146:531-539.
52. Arican O, Aral M, Sasmaz S, Ciragil P. Serum levels of TNF-α, IFN-γ, IL-6, IL-8, IL-12, IL-17, and IL-18 in patients with active psoriasis and correlation with disease severity. Mediators Inflamm 2005; 2005:273-279.
53. Demircan N, Safran B, Soylu M, Ozcan A, Sizmaz S. Determination of vitreous interleukin-1 (IL-1) and tumour necrosis factor (TNF) levels in proliferative diabetic retinopathy. Eye 2006; 20:1366-1369.
54. Shi Y-Q, Huang T-W, Chen L-M, Pan X-D, Zhang J, Zhu Y-G, et al. Ginsenoside Rg1 attenuates amyloid-β content, regulates PKA/CREB activity, and improves cognitive performance in SAMP8 mice. J Alzheimers Dis 2010; 19:977-989.
55. Lee B-H, Kim H, Park S-H, Kim Y-K. Decreased plasma BDNF level in depressive patients. J Affect Disord 2007; 101:239-244.
56. Robinson DS, Gilmor ML, Yang Y, Moonsammy G, Azzaro AJ, Oren DA, et al. Treatment effects of selegiline transdermal system on symptoms of major depressive disorder: a meta-analysis of short-term, placebo-controlled, efficacy trials. Psychopharmacol Bull 2007; 40:15-28.
57. Veazey C, Ozlem Erden Aki S, Cook KF, Lai EC, Kunik ME. Prevalence and treatment of depression in Parkinson’s disease. J Neuropsychiatry Clin Neurosci 2005; 17:310-323.
58. Yue F, Zeng S, Wu D, Yi D, Zhang YA, Chan P. Age-related decline in motor behavior and striatal dopamine transporter in cynomolgus monkeys. J Neural Transm 2012; 119:943-952.
59. R-ORAL A. Neurobiology of neuro-psychiatric diseases. J Neural Transm 2008; 115:1463-1483.
60. Volkow ND, Fowler JS, Wang G-J, Swanson JM, Telang F. Dopamine in drug abuse and addiction: results of imaging studies and treatment implications.  Arch Neurol 2007; 64:1575-1579.
61. Hamaue N, Minami M, Terado M, Hirafuji M, Endo T, Machida M, et al. Comparative study of the effects of isatin, an endogenous MAO-inhibitor, and selegiline on bradykinesia and dopamine levels in a rat model of Parkinson’s disease induced by the Japanese encephalitis virus. Neurotoxicology 2004; 25:205-213.
62. Halladay AK, Kusnecov A, Michna L, Kita T, Hara C, Wagner GC. Relationship between methamphetamine‐induced dopamine release, hyperthermia, self‐injurious behaviour and long term dopamine depletion in BALB/c and C57BL/6 mice. Pharmacol Toxicol 2003; 93:33-41.
63. Dalal A, Poddar MK. Short-term erythrosine B-induced inhibition of the brain regional serotonergic activity suppresses motor activity (exploratory behavior) of young adult mammals. Pharmacol Biochem Behav 2009; 92:574-582.
64. Cao G, Zhu J, Zhong Q, Shi C, Dang Y, Han W, et al. Distinct roles of methamphetamine in modulating spatial memory consolidation, retrieval, reconsolidation and the accompanying changes of ERK and CREB activation in hippocampus and prefrontal cortex. Neuropharmacology 2013; 67:144-154.
65. Goverdhan P, Sravanthi A, Mamatha T. Neuroprotective effects of meloxicam and selegiline in scopolamine-induced cognitive impairment and oxidative stress. Int J Alzheimers Dis 2012; 2012.94013.
66. West BD, Shughrue PJ, Vanko AE, Ransom RW, Kinney GG. Amphetamine-induced locomotor activity is reduced in mice following MPTP treatment but not following selegiline/MPTP treatment. Pharmacol Biochem Behav 2006; 84:158-161.
67. Bert B, Harms S, Langen B, Fink H. Clomipramine and selegiline: do they influence impulse control? J Vet Pharmacol Ther 2006; 29:41-47.
68. LaVoie MJ, Card JP, Hastings TG. Microglial activation precedes dopamine terminal pathology in methamphetamine-induced neurotoxicity. Exp Neurol 2004; 187:47-57.
69. Riddle EL, Fleckenstein AE, Hanson GR. Mechanisms of methamphetamine-induced dopaminergic neurotoxicity. AAPS J 2006; 8:E413-E418.
70. Melo P, Rodrigues LG, Pinazo‐Durán MD, Tavares MA. Methamphetamine and lipid peroxidation in the rat retina. Birth Defects Res A Clin Mol Teratol 2005; 73:455-460.
71. Zhu W, Xie W, Pan T, Jankovic J, Li J, Youdim MB, et al. Comparison of neuroprotective and neurorestorative capabilities of rasagiline and selegiline against lactacystin‐induced nigrostriatal dopaminergic degeneration. J Neurochem 2008; 105:1970-1978.
72. McKinley ET, Baranowski TC, Blavo DO, Cato C, Doan TN, Rubinstein AL. Neuroprotection of MPTP-induced toxicity in zebrafish dopaminergic neurons. Mol Brain Res 2005; 141:128-137.
73. Szökő É, Tábi T, Riederer P, Vécsei L, Magyar K. Pharmacological aspects of the neuroprotective effects of irreversible MAO-B inhibitors, selegiline and rasagiline, in Parkinson’s disease. J Neural Transm (Vienna) 2018; 121:1-15.
74. Mannerström M, Toimela T, Ylikomi T, Tähti H. The combined use of human neural and liver cell lines and mouse hepatocytes improves the predictability of the neurotoxicity of selected drugs. Toxicol Lett 2006; 165:195-202.
75. Krasnova IN, Chiflikyan M, Justinova Z, McCoy MT, Ladenheim B, Jayanthi S, et al. CREB phosphorylation regulates striatal transcriptional responses in the self-administration model of methamphetamine addiction in the rat. Neurobiol Dis 2013; 58:132-143.
76. Krasnova IN, Justinova Z, Cadet JL. Methamphetamine addiction: involvement of CREB and neuroinflammatory signaling pathways. Psychopharmacology (Berl) 2016; 233:1945-1962.
77. Motaghinejad M, Motevalian M, Fatima S, Beiranvand T, Mozaffari S. Topiramate via NMDA, AMPA/kainate, GABAA and Alpha2 receptors and by modulation of CREB/BDNF and Akt/GSK3 signaling pathway exerts neuroprotective effects against methylphenidate-induced neurotoxicity in rats. J J Neural Transm (Vienna) 2017;124:1369-1387
78. Motaghinejad M, Motevalian M, Abdollahi M, Heidari M, Madjd Z. Topiramate confers neuroprotection against methylphenidate-induced neurodegeneration in dentate gyrus and CA1 regions of Hippocampus via CREB/BDNF pathway in rats. Neurotox Res 2017; 31:373-399.
79. Naoi M, Maruyama W, Inaba-Hasegawa K. Revelation in the neuroprotective functions of rasagiline and selegiline: the induction of distinct genes by different mechanisms. Expert Rev Neurother 2013; 13:671-684.