Neurotoxicity Research

, Volume 35, Issue 1, pp 150–159 | Cite as

The Antidiabetic Drug Liraglutide Minimizes the Non-Cholinergic Neurotoxicity of the Pesticide Mipafox in SH-SY5Y Cells

  • Laís Silva Fernandes
  • Neife Aparecida G. dos Santos
  • Guilherme Luz Emerick
  • Antonio Cardozo dos SantosEmail author


Organophosphorus (OPs) compounds have been widely used in agriculture, industry, and household, and the neurotoxicity induced by them is still a cause of concern. The main toxic mechanism of OPs is the inhibition of acetylcholinesterase (AChE); however, the delayed neuropathy induced by OPs (OPIDN) is mediated by other mechanisms such as the irreversible inhibition of 70% of NTE activity (neuropathy target esterase) that leads to axonal degeneration. Liraglutide is a long-lasting GLP-1 analog clinically used as antidiabetic. Its neurotrophic and neuroprotective effects have been demonstrated in vitro and in experimental models of neurodegenerative diseases. As in OPIDN, axonal degeneration also plays a role in neurodegenerative diseases. Therefore, this study investigated the protective potential of liraglutide against the neurotoxicity of OPs by using mipafox as a neuropathic agent (at a concentration able to inhibit and age 70% of NTE activity) and a neuronal model with SH-SY5Y neuroblastoma cells, which express both esterases. Liraglutide protected cells against the neurotoxicity of mipafox by increasing neuritogenesis, the uptake of glucose, the levels of cytoskeleton proteins, and synaptic-plasticity modulators, besides decreasing the pro-inflammatory cytokine interleukin 1β and caspase-3 activity. This is the first study to suggest that liraglutide might induce beneficial effects against the delayed, non-cholinergic neurotoxicity of OPs.


Liraglutide Mipafox Organophosphate-induced delayed neuropathy (OPIDN) Neuroplasticity Neuroprotection 


Funding Information

This study received financial support from Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, Grants 307657/2016-7 and 140105/2015-8) and Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP, Grants 2012/00168-6, 2012/16319-3 and 2013/26906-6).

Compliance with Ethical Standards

Conflict of Interest Statement

The authors declare that they have no conflicts of interest.


  1. Abdollahi M, Donyavi M, Pournourmohammadi S, Saadat M (2004) Hyperglycemia associated with increased hepatic glycogen phosphorylase and phosphoenolpyruvate carboxykinase in rats following subchronic exposure to malathion. Comp Biochem Physiol C Toxicol Pharmacol 137:343–347CrossRefGoogle Scholar
  2. Abou-Donia MB (1993) The cytoskeleton as a target for organophosphorus ester-induced delayed neurotoxicity (OPIDN). Chem Biol Interact 87:383–393CrossRefGoogle Scholar
  3. Andreozzi F, Raciti GA, Nigro C, Mannino GC, Procopio T, Davalli AM, Beguinot F, Sesti G, Miele C, Folli F (2016) The GLP-1 receptor agonists exenatide and liraglutide activate glucose transport by an AMPK-dependent mechanism. J Transl Med 14:229CrossRefGoogle Scholar
  4. Androutsopoulos VP, Hernandez AF, Liesivuori J, Tsatsakis AM (2013) A mechanistic overview of health associated effects of low levels of organochlorine and organophosphorous pesticides. Toxicology 307:89–94CrossRefGoogle Scholar
  5. Arsenault AL, Gibson MA, Mader ME (1975) Hypoglycemia in malathion-treated chick embryos. Can J Zool 53:1055–1057CrossRefGoogle Scholar
  6. Athamneh AIM, He Y, Lamoureux P, Fix L, Suter DM, Miller KE (2017) Neurite elongation is highly correlated with bulk forward translocation of microtubules. Sci Rep 7:7292CrossRefGoogle Scholar
  7. Athauda D, Foltynie T (2016) The glucagon-like peptide 1 (GLP) receptor as a therapeutic target in Parkinson’s disease: mechanisms of action. Drug Discov Today 21:802–818CrossRefGoogle Scholar
  8. Banks CN, Lein PJ (2012) A review of experimental evidence linking neurotoxic organophosphorus compounds and inflammation. Neurotoxicology 33:575–584CrossRefGoogle Scholar
  9. Bauer S, Kerr BJ, Patterson PH (2007) The neuropoietic cytokine family in development, plasticity, disease and injury. Nat Rev Neurosci 8:221–232CrossRefGoogle Scholar
  10. Benomar Y, Naour N, Aubourg A, Bailleux V, Gertler A, Djiane J, Guerre-Millo ML, Taouis M (2006) Insulin and leptin induce Glut4 plasma membrane translocation and glucose uptake in a human neuronal cell line by a phosphatidylinositol 3-kinase- dependent mechanism. Endocrinology 147:2550–2556CrossRefGoogle Scholar
  11. Bissonnette CJ, Klegeris A, Mcgeer PL, Mcgeer EG (2004) Interleukin 1alpha and interleukin 6 protect human neuronal SH-SY5Y cells from oxidative damage. Neurosci Lett 361:40–43CrossRefGoogle Scholar
  12. Burnette WN (1981) "Western blotting." Electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem 112:195–203CrossRefGoogle Scholar
  13. Carlson K, Ehrich M (2001) Organophosphorus compounds alter intracellular F-actin content in SH-SY5Y human neuroblastoma cells. Neurotoxicology 22:819–827CrossRefGoogle Scholar
  14. Caudle WM (2015) Chapter 14 - Occupational exposures and parkinsonism. In: Lotti M, Bleecker ML (eds) Handbook of clinical neurology. ElsevierGoogle Scholar
  15. Chang PA, Wu YJ (2006) Effect of tri-o-cresyl phosphate on cytoskeleton in human neuroblastoma SK-N-SH cell. Mol Cell Biochem 290:145–151CrossRefGoogle Scholar
  16. Chen LN, Lyu J, Yang XF, Ji WJ, Yuan BX, Chen MX, Ma X, Wang B (2013) Liraglutide ameliorates glycometabolism and insulin resistance through the upregulation of GLUT4 in diabetic KKAy mice. Int J Mol Med 32:892–900CrossRefGoogle Scholar
  17. CHO T, Tiffany-Castiglioni E (2004) Neurofilament 200 as an indicator of differences between mipafox and paraoxon sensitivity in Sy5Y neuroblastoma cells. J Toxicol Environ Health A 67:987–1000CrossRefGoogle Scholar
  18. Cogulu D, Onay H, Ozdemir Y, Aslan GI, Ozkinay F, Kutukculer N, Eronat C (2015) Associations of interleukin (IL)-1beta, IL-1 receptor antagonist, and IL-10 with dental caries. J Oral Sci 57:31–36CrossRefGoogle Scholar
  19. Das KP, Freudenrich TM, Mundy WR (2004) Assessment of PC12 cell differentiation and neurite growth: a comparison of morphological and neurochemical measures. Neurotoxicol Teratol 26:397–406CrossRefGoogle Scholar
  20. Ehrich M, Jortner BS (2001) Organophosphate-induced delayed neuropathy. In: Massaro EJ (ed) Handbook of Neurotoxicology. Humana Press Inc., TotowaGoogle Scholar
  21. Ehrich M, Correll L, Veronesi B (1997) Acetylcholinesterase and neuropathy target esterase inhibitions in neuroblastoma cells to distinguish organophosphorus compounds causing acute and delayed neurotoxicity. Fundam Appl Toxicol 38:55–63CrossRefGoogle Scholar
  22. el-Fawal HA, Correll L, Gay L, Ehrich M (1990) Protease activity in brain, nerve, and muscle of hens given neuropathy-inducing organophosphates and a calcium channel blocker. Toxicol Appl Pharmacol 103:133–142CrossRefGoogle Scholar
  23. Emerick GL, Deoliveira GH, dos Santos AC, Ehrich M (2012) Mechanisms for consideration for intervention in the development of organophosphorus-induced delayed neuropathy. Chem Biol Interact 199:177–184CrossRefGoogle Scholar
  24. Emerick GL, Fernandes LS, de Paula ES, Barbosa F Jr, dos Santos NA, dos Santos AC (2015) In vitro study of the neuropathic potential of the organophosphorus compounds fenamiphos and profenofos: comparison with mipafox and paraoxon. Toxicol in Vitro 29:1079–1087CrossRefGoogle Scholar
  25. Erta M, Quintana A, Hidalgo J (2012) Interleukin-6, a major cytokine in the central nervous system. Int J Biol Sci 8:1254–1266CrossRefGoogle Scholar
  26. Fernandes LS, Emerick GL, dos Santos NA, de Paula ES, Barbosa F Jr, dos Santos AC (2015) In vitro study of the neuropathic potential of the organophosphorus compounds trichlorfon and acephate. Toxicol in Vitro 29:522–528CrossRefGoogle Scholar
  27. Fernandes LS, dos Santos NAG, Emerick GL, Santos ACD (2017) L- and T-type calcium channel blockers protect against the inhibitory effects of mipafox on neurite outgrowth and plasticity-related proteins in SH-SY5Y cells. J Toxicol Environ Health A 80:1086–1097CrossRefGoogle Scholar
  28. Gill KD, Flora G, Pachauri V, Flora SJS (2010) Neurotoxicity of organophosphates and carbamates. In: Satoh T, Gupta RC (eds) Anticholinesterase pesticides. Metabolism, neurotoxicity, and epidemiology. Wiley, New JerseyGoogle Scholar
  29. Hans VH, Kossmann T, Lenzlinger PM, Probstmeier R, Imhof HG, Trentz O, Morganti-Kossmann MC (1999) Experimental axonal injury triggers interleukin-6 mRNA, protein synthesis and release into cerebrospinal fluid. J Cereb Blood Flow Metab 19:184–194CrossRefGoogle Scholar
  30. Husain K (2014) Delayed neurotoxicity of organophosphorus Compounds. J Environ Immunol Toxicol 1:7Google Scholar
  31. Jiang Y, Liu X, Li S, Zhang Y, piao F, Sun X (2014) Identification of differentially expressed proteins related to organophosphorus-induced delayed neuropathy in the brains of hens. J Appl Toxicol 34:1352–1360CrossRefGoogle Scholar
  32. Johnson MK (1974) The primary biochemical lesion leading to the delayed neurotoxic effects of some organophosphorus esters. J Neurochem 23:785–789CrossRefGoogle Scholar
  33. Johnson MK (1982) The target site for the initiation of delayed neurotoxicity by organophosphorous esters: biochemical studies and toxicological applications. In: Hodgson JR, Philpot RM (eds) Rev Biochem ToxicolGoogle Scholar
  34. Kanaan NM, Pigino GF, Brady ST, Lazarov O, Binder LI, Morfini GA (2013) Axonal degeneration in Alzheimer’s disease: when signaling abnormalities meet the axonal transport system. Exp Neurol 246:44–53CrossRefGoogle Scholar
  35. Karami-Mohajeri S, Abdollahi M (2011) Toxic influence of organophosphate, carbamate, and organochlorine pesticides on cellular metabolism of lipids, proteins, and carbohydrates: a systematic review. Hum Exp Toxicol 30:1119–1140CrossRefGoogle Scholar
  36. Kevenaar JT, Hoogenraad CC (2015) The axonal cytoskeleton: from organization to function. Front Mol Neurosci 8Google Scholar
  37. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685CrossRefGoogle Scholar
  38. L'episcopo F, Serapide MF, Tirolo C, Testa N, Caniglia S, Morale MC, Pluchino S, Marchetti B (2011) A Wnt1 regulated frizzled-1/beta-catenin signaling pathway as a candidate regulatory circuit controlling mesencephalic dopaminergic neuron-astrocyte crosstalk: therapeutical relevance for neuron survival and neuroprotection. Mol Neurodegener 6:49CrossRefGoogle Scholar
  39. Li Z, Ni CL, Yao Z, Chen LM, Niu WY (2014) Liraglutide enhances glucose transporter 4 translocation via regulation of AMP-activated protein kinase signaling pathways in mouse skeletal muscle cells. Metabolism 63:1022–1030CrossRefGoogle Scholar
  40. Li M, Li S, Li Y (2015a) Liraglutide promotes cortical neurite outgrowth via the MEK-ERK pathway. Cell Mol Neurobiol 35:987–993CrossRefGoogle Scholar
  41. Li Y, Bader M, Tamargo I, Rubovitch V, Tweedie D, Pick CG, Greig NH (2015b) Liraglutide is neurotrophic and neuroprotective in neuronal cultures and mitigates mild traumatic brain injury in mice. J Neurochem 135:1203–1217CrossRefGoogle Scholar
  42. Long-Smith CM, Manning S, McClean PL, Coakley MF, O'Halloran DJ, Holscher C, O'Neill C (2013) The diabetes drug liraglutide ameliorates aberrant insulin receptor localisation and signalling in parallel with decreasing both amyloid-beta plaque and glial pathology in a mouse model of Alzheimer’s disease. NeuroMolecular Med 15:102–114CrossRefGoogle Scholar
  43. Lotti M (1991) The pathogenesis of organophosphate polyneuropathy. Crit Rev Toxicol 21:465–487CrossRefGoogle Scholar
  44. Lotti M, Moretto A, Capodicasa E, Bertolazzi M, Peraica M, Scapellato ML (1993) Interactions between neuropathy target esterase and its inhibitors and the development of polyneuropathy. Toxicol Appl Pharmacol 122:165–171CrossRefGoogle Scholar
  45. Mahmood T, Yang PC (2012) Western blot: technique, theory, and trouble shooting. N Am J Med Sci 4:429–434CrossRefGoogle Scholar
  46. Mainardi M, Fusco S, Grassi C (2015) Modulation of hippocampal neural plasticity by glucose-related signaling. Neural Plast 2015:657928CrossRefGoogle Scholar
  47. McClean PL, Parthsarathy V, Faivre E, Holscher C (2011) The diabetes drug liraglutide prevents degenerative processes in a mouse model of Alzheimer’s disease. J Neurosci 31:6587–6594CrossRefGoogle Scholar
  48. Meredith C, Johnson MK (1988) Neuropathy target esterase: rates of turnover in vivo following covalent inhibition with phenyl di-n-pentylphosphinate. J Neurochem 51:1097–1101CrossRefGoogle Scholar
  49. Murphy PG, Grondin J, Altares M, Richardson PM (1995) Induction of interleukin-6 in axotomized sensory neurons. J Neurosci 15:5130–5138CrossRefGoogle Scholar
  50. Narayan S, Liew Z, Paul K, Lee P-C, Sinsheimer JS, Bronstein JM, Ritz B (2013) Household organophosphorus pesticide use and Parkinson’s disease. Int J Epidemiol 42:1476–1485CrossRefGoogle Scholar
  51. Parthsarathy V, Holscher C (2013a) Chronic treatment with the GLP1 analogue liraglutide increases cell proliferation and differentiation into neurons in an AD mouse model. PLoS One 8:e58784CrossRefGoogle Scholar
  52. Perry T, Lahiri DK, Chen D, Zhou J, Shaw KT, Egan JM, Greig NH (2002) A novel neurotrophic property of glucagon-like peptide 1: a promoter of nerve growth factor-mediated differentiation in PC12 cells. J Pharmacol Exp Ther 300:958–966CrossRefGoogle Scholar
  53. Porseva VV, Smirnova VP, Korzina MB, Emanuilov AI, Masliukov PM (2013) Age-associated changes in sympathetic neurons containing neurofilament 200 kDa during chemical deafferentation. Bull Exp Biol Med 155:268–271CrossRefGoogle Scholar
  54. Postuma RB, Saez-Valero J, Small DH (1999) Inhibition of neurite outgrowth from chick sympathetic neurons by cholinesterase inhibitors is not mediated by binding to cholinesterases. Neurosci Lett 266:77–80CrossRefGoogle Scholar
  55. Pournourmohammadi S, Farzami B, Ostad SN, Azizi E, Abdollahi M (2005) Effects of malathion subchronic exposure on rat skeletal muscle glucose metabolism. Environ Toxicol Pharmacol 19:191–196CrossRefGoogle Scholar
  56. Raghunath M, Patti R, Bannerman P, Lee CM, Baker S, Sutton LN, Phillips PC, Damodar Reddy C (2000) A novel kinase, AATYK induces and promotes neuronal differentiation in a human neuroblastoma (SH-SY5Y) cell line. Brain Res Mol Brain Res 77:151–162CrossRefGoogle Scholar
  57. Rasband WS 1997-2014. ImageJ, U. S. National Institutes of Health. Bethesda, Maryland, USA
  58. Ren K, Torres R (2009) Role of interleukin-1β during pain and inflammation. Brain Res Rev 60:57–64CrossRefGoogle Scholar
  59. Richardson RJ, Worden RM, Wijeyesakere SJ, Hein ND, Fink JK, Makhaeva GF (2015) Chapter 63 - Neuropathy target esterase as a biomarker and biosensor of delayed neuropathic agents A2. In: Gupta RC (ed) Handbook of toxicology of chemical warfare agents, 2nd edn. Academic Press, BostonGoogle Scholar
  60. Rohlman DS, Anger WK, Lein PJ (2011) Correlating neurobehavioral performance with biomarkers of organophosphorous pesticide exposure. Neurotoxicology 32:268–276CrossRefGoogle Scholar
  61. Romero-Navarro G, Lopez-Aceves T, Rojas-Ochoa A, Fernandez Mejia C (2006) Effect of dichlorvos on hepatic and pancreatic glucokinase activity and gene expression, and on insulin mRNA levels. Life Sci 78:1015–1020CrossRefGoogle Scholar
  62. Russo VC, Kobayashi K, Najdovska S, Baker NL, Werther GA (2004) Neuronal protection from glucose deprivation via modulation of glucose transport and inhibition of apoptosis: a role for the insulin-like growth factor system. Brain Res 1009:40–53CrossRefGoogle Scholar
  63. Sachana M, Flaskos J, Alexaki E, Hargreaves AJ (2003) Inhibition of neurite outgrowth in N2a cells by leptophos and carbaryl: effects on neurofilament heavy chain, GAP-43 and HSP-70. Toxicol in Vitro 17:115–120CrossRefGoogle Scholar
  64. Scheller J, Chalaris A, Schmidt-Arras D, Rose-John S (2011) The pro- and anti-inflammatory properties of the cytokine interleukin-6. Biochim Biophys Acta 1813:878–888CrossRefGoogle Scholar
  65. Schimmelpfeng J, Weibezahn KF, Dertinger H (2004) Quantification of NGF-dependent neuronal differentiation of PC-12 cells by means of neurofilament-L mRNA expression and neuronal outgrowth. J Neurosci Methods 139:299–306CrossRefGoogle Scholar
  66. Schneider L, Giordano S, Zelickson BR, Johnson SM, Bernavides AG, Ouyang X, Fineberg N, Darley-Usmar VM, Zhang J (2011) Differentiation of SH-SY5Y cells to a neuronal phenotype changes cellular bioenergetics and the response to oxidative stress. Free Radic Biol Med 51:2007–2017CrossRefGoogle Scholar
  67. Sharma MK, Jalewa J, Holscher C (2014) Neuroprotective and anti-apoptotic effects of liraglutide on SH-SY5Y cells exposed to methylglyoxal stress. J Neurochem 128:459–471CrossRefGoogle Scholar
  68. Sharma AN, Ligade SS, Sharma JN, Shukla P, Elased KM, Lucot JB (2015) GLP-1 receptor agonist liraglutide reverses long-term atypical antipsychotic treatment associated behavioral depression and metabolic abnormalities in rats. Metab Brain Dis 30:519–527CrossRefGoogle Scholar
  69. Tagliaferro P, Burke RE (2016) Retrograde axonal degeneration in Parkinson disease. J Parkinsons Dis 6:1–15CrossRefGoogle Scholar
  70. Tagliati M (2017) A phase II, randomized, double-blinded, placebo-controlled trial of liraglutide in Parkinson’s disease [Online].
  71. Terry AV Jr (2012) Functional consequences of repeated organophosphate exposure: potential non-cholinergic mechanisms. Pharmacol Ther 134:355–365CrossRefGoogle Scholar
  72. Thorens B, Mueckler M (2010) Glucose transporters in the 21st century. Am J Physiol Endocrinol Metab 298:E141–E145CrossRefGoogle Scholar
  73. Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A 76:4350–4354CrossRefGoogle Scholar
  74. Vester A, Caudle WM (2016) The synapse as a central target for neurodevelopmental susceptibility to pesticides. Toxics 4Google Scholar
  75. Voorhees JR, Rohlman DS, Lein PJ, Pieper AA (2017) Neurotoxicity in preclinical models of occupational exposure to organophosphorus compounds. Front Neurosci 10Google Scholar
  76. Xiong H, Zheng C, Wang J, Song J, Zhao G, Shen H, Deng Y (2013) The neuroprotection of liraglutide on Alzheimer-like learning and memory impairment by modulating the hyperphosphorylation of tau and neurofilament proteins and insulin signaling pathways in mice. J Alzheimers Dis 37:623–635CrossRefGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Laís Silva Fernandes
    • 1
  • Neife Aparecida G. dos Santos
    • 1
  • Guilherme Luz Emerick
    • 2
  • Antonio Cardozo dos Santos
    • 1
    Email author
  1. 1.Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto—FCFRPUniversidade de São Paulo, USPRibeirão PretoBrazil
  2. 2.Instituto de Ciências da SaúdeUniversidade Federal de Mato Grosso—ICS/UFMT/CUSSinopBrazil

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