Cannabinoid Receptor Type 1 Agonist ACEA Protects Neurons from Death and Attenuates Endoplasmic Reticulum Stress-Related Apoptotic Pathway Signaling

Abstract

Neurodegeneration is the result of progressive destruction of neurons in the central nervous system, with unknown causes and pathological mechanisms not yet fully elucidated. Several factors contribute to neurodegenerative processes, including neuroinflammation, accumulation of neurotoxic factors, and misfolded proteins in the lumen of the endoplasmic reticulum (ER). Endocannabinoid signaling has been pointed out as an important modulatory system in several neurodegeneration-related processes, inhibiting the inflammatory response and increasing neuronal survival. Thus, we investigated the presumptive protective effect of the selective cannabinoid type 1 (CB1) receptor agonist arachidonyl-2′-chloroethylamide (ACEA) against inflammatory (lipopolysaccharide, LPS) and ER stress (tunicamycin) stimuli in an in vitro neuronal model (Neuro-2a neuroblastoma cells). Cell viability analysis revealed that ACEA was able to protect against cell death induced by LPS and tunicamycin. This neuroprotective effect occurs via the CB1 receptor in the inflammation process and via the transient receptor potential of vanilloid type-1 (TRPV1) channel in ER stress. Furthermore, the immunoblotting analyses indicated that the neuroprotective effect of ACEA seems to involve the modulation of eukaryotic initiation factor 2 (eIF2α), transcription factor C/EBP homologous protein (CHOP), and caspase 12, as well as the survival/death p44/42 MAPK, ERK1/2-related signaling pathways. Together, these data suggest that the endocannabinoid system is a potential therapeutic target in neurodegenerative processes, especially in ER-related neurodegenerative diseases.

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References

  1. Aso E, Ferrer I (2014) Cannabinoids for treatment of Alzheimer’s disease: moving toward the clinic. Front Pharmacol 5:10–19

    Article  Google Scholar 

  2. Batinga H, Zúñiga-Hertz JP, Torrão AS (2016) Cannabinoid receptor ligands prevent dopaminergic neurons death induced by neurotoxic, inflammatory and oxidative stimuli in vitro. J Biomed Sci 5:1–16

    Article  Google Scholar 

  3. Benito C, Romero JP, Tolón RM, Clemente D, Docagne F, Hillard CJ, Guaza C, Romero J (2007) Cannabinoid CB1 and CB2 receptors and fatty acid amide hydrolase are specific markers of plaque cell subtypes in human multiple sclerosis. J Neurosci 27:2396–2402

    CAS  Article  PubMed  Google Scholar 

  4. Bitko V, Barik S (2001) An endoplasmic reticulum-specific stress-activated caspase (caspase-12) is implicated in the apoptosis of A549 epithelial cells by respiratory syncytial virus. J Cell Biochem 80:441–454

    CAS  Article  PubMed  Google Scholar 

  5. Bouaboula M, Poinot-Chazel C, Bourrié B, Canat X, Calandra B, Rinaldi-Carmona M, Le Fur G, Casellas P (1995) Activation of mitogen-activated protein kinases by stimulation of the central cannabinoid receptor CB1. Biochem J 312:637–641

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  6. Caballero B, Coto-Montes A (2012) An insight into the role of autophagy in cell responses in the aging and neurodegenerative brain. Histol Histopathol 27:263–275

    CAS  PubMed  Google Scholar 

  7. Café-Mendes CC, Ferro ES, Torrão AS, Crunfli F, Rioli V, Schmitt A, Falkai P, Britto LR, Turck CW, Martins-de-Souza D (2017) Peptidomic analysis of the anterior temporal lobe and corpus callosum from schizophrenia patients. J Proteome 151:97–105

    Article  Google Scholar 

  8. Calderón FH, Bonnefont A, Muñoz FJ, Fernández V, Videla LA, Inestrosa NC (1999) PC12 and neuro 2ª cells have diferente susceptibilities to acetylcholinesterase-amyloid complexes, amyloid25-35 fragnebr, glutamate, and hydrogen peroxide. J Neurosci Res 56:620–631

    Article  PubMed  Google Scholar 

  9. Caltana LM, Heimrich B, Brusco A (2015a) Further evidence for the neuroplastic role of cannabinoids: a study in organotypic hippocampal slice cultures. J Mol Neurosci 56:773–781

    CAS  Article  PubMed  Google Scholar 

  10. Caltana L, Saez TM, Aronne MP, Brusco A (2015b) Cannabinoid receptor type 1 agonist ACEA improves motor recovery and protects neurons in ischemic stroke in mice. J Neurochem 135:616–629

    CAS  Article  PubMed  Google Scholar 

  11. Chung YC, Bok E, Huh SH, Park JY, Yoon SH, Kim SR, Kim YS, Maeng S, Park SH, Jin BK (2011) Cannabinoid receptor type 1 protects nigrostriatal dopaminergic neurons against MPTP neurotoxicity by inhibiting microglial activation. J Immunol 187:6508–6517

    CAS  Article  PubMed  Google Scholar 

  12. Croxford JL, Yamamura T (2005) Cannabinoids and the immune system: potential for the treatment of inflammatory diseases? J Neuroimmun 166:3–18

    CAS  Article  Google Scholar 

  13. De Mario A, Quintana-Cabrera R, Martinvalet D, Giacomello M (2017) (Neuro) degenerated mitochondria-ER contacts Biochem. Biophys Res Commun 483:1096–1109

    Article  Google Scholar 

  14. Di Marzo V (2008) Targeting the endocannabinoid system: to enhance or reduce? Nat Rev Drug Discov 7:338–455

    Article  Google Scholar 

  15. Di Marzo V, De Petrocellis L (2012) Why do cannabinoid receptors have more than one endogenous ligand? Philos Trans R SocLond B BiolSci 367:3216–3228

    Article  Google Scholar 

  16. Fernández-Ruiz J, Romero J, Ramos JA (2015) Endocannabinoids and neurodegenerative disorders: Parkinson’s disease, Huntington’s chorea, Alzheimer’s disease, and others. Handb Exp Pharmacol 231:233–259

    Article  PubMed  Google Scholar 

  17. Forman MS, Lee VM, Trojanowsi JQ (2003) “Unfolding” pathways in neurodegenerative disease. Trends Neurosci 26:407–410

    CAS  Article  PubMed  Google Scholar 

  18. Fowler CJ, Rojo ML, Rodriguez-Gaztelumendi A (2010) Modulation of the endocannabinoid system: neuroprotective or neurotoxicity? Exp Neurol 224:37–47

    CAS  Article  PubMed  Google Scholar 

  19. Frank-Cannon TC, Alto LT, McAlpine FE, Tansey MG (2009) Does neuroinflammation fan the flame in neurodegenerative diseases? Bio Med Central 4:40–47

    Google Scholar 

  20. Galve-Roperh I, Rueda D, Pulgar TG, Velasco G, Guzmán M (2002) Mechanism of extracellular signal-regulated kinase activation by the CB1 cannabinoid receptor. Mol Pharmacol 62:1385–1392

    CAS  Article  PubMed  Google Scholar 

  21. Gilbert GL, Kim HJ, Waataja JJ, Thayer SA (2007) Delta (9)-tetrahydrocannabinol protects hippocampal neurons from excitotoxicity. Brain Res 1128:61–69

    CAS  Article  PubMed  Google Scholar 

  22. Grewal SS, York RD, Stork PJS (1999) Extracellular-signal-regulated kinase signaling in neurons. Curr Opin Neurobiol 9:544–553

    CAS  Article  PubMed  Google Scholar 

  23. Glenner GG, Wong CW (1984) Alzheimer’s disease: initial report of the purification and characterization of a novel cerebrovascular amyloid protein. Biochem Biophys Res Commun 120:885–890

    CAS  Article  PubMed  Google Scholar 

  24. Grundy RI, Rabuffetti M, Beltramo M (2001) Cannabinoids and neuroprotection. Mol Neurobiol 24:29–51

    CAS  Article  PubMed  Google Scholar 

  25. Guzmán M (2003) Neurons on cannabinoids: dead or alive? Br J Pharmacol 140:439–440

    Article  PubMed  PubMed Central  Google Scholar 

  26. Harvey BS, Ohlsson KS, Maag JL, Musgrave IF, Smid SD (2012) Contrasting protective effects of cannabinoids against oxidative stress and amyloid-β evoked neurotoxicity in vitro. Neurotoxicology 33:138–146

    CAS  Article  PubMed  Google Scholar 

  27. Hetz C, Mollereau B (2014) Disturbance of endoplasmic reticulum proteostasis in neurodegenerative diseases. Nat Rev Neurosci 15:233–249

    CAS  Article  PubMed  Google Scholar 

  28. Hillard CJ, Manna S, Greenberg MJ, DiCamelli R, Ross RA, Stevenson LA, Murphy V, Pertwee RG, Campbell WB (1999) Synthesis and characterization of potent and selective agonists of the neuronal cannabinoid receptor (CB1). J Pharmacol Exp Ther 289:1427–1433

    CAS  PubMed  Google Scholar 

  29. Hoozemans JJ, Van Haastert ES, Nijholt DA, Rozemuller AJ, Scheper W (2012) Activation of the unfolded protein response is an early event in Alzheimer’s and Parkinson’s disease. Neurodegener Dis 10:212–215

    CAS  Article  PubMed  Google Scholar 

  30. Huang CC, Faber PW, Persichetti F, Mittal V, Vonsattel JP, Macdonald ME, Gusella JF (1998) Amyloid formation by mutant huntingtin: threshold, progressivity and recruitment of normal polyglutamine proteins. Somat Cell Mol Genet 24:217–233

    CAS  Article  PubMed  Google Scholar 

  31. Jordan JD, He JC, Eungdamrong NJ, Gomes I, Ali W, Nguyen T, Bivona TG, Philips MR, Devi LA, Iyengar R (2005) Cannabinoid receptor-induced neurite outgrowth is mediated by Rap1 activation through G(alpha)o/i-triggered proteasomal degradation of Rap1GAPII. J Biol Chem 280:11413–11421

    CAS  Article  PubMed  Google Scholar 

  32. Lan R, Liu Q, Fan P, Lin S, Fernando SR, McCallion D, Pertwee R, Makriyannis A (1999) Structure-activity relationships of pyrazolederivates as cannabinoid receptor antagonists. J Med Chem 42:769–776

    CAS  Article  PubMed  Google Scholar 

  33. Lastres-Becker I, Molina-Holgado F, Ramos JA, Mechoulam R, Fernández-Ruiz J (2005) Cannabinoids provide neuroprotection against 6-hydroxydopamine toxicity in vivo and in vitro: relevance to Parkinson’s disease. Neurobiol Dis 19:96–107

    CAS  Article  PubMed  Google Scholar 

  34. Lisboa SF, Guimarães FS (2012) Differential role of CB1 and TRPV1 receptors on anandamide modulation of defensive responses induced by nitric oxide in the dorsolateral periaqueductal gray. Neuropharmacology 62:2455–2462

    CAS  Article  PubMed  Google Scholar 

  35. Ma Y, Brewer JW, Diehl JA, Hendershot LM (2002) Two distinct stress signaling pathways converge upon the CHOP promoter during the mammalian unfolded protein response. J Mol Biol 318:1351–1365

    CAS  Article  PubMed  Google Scholar 

  36. Ma L, Jia J, Niu W, Jiang T, Zhai Q, Yang L, Bai F, Wang Q, Xiong L (2015) Mitochondrial CB1 receptor is involved in ACEA-induced protective effects on neurons and mitochondrial functions. Sci Rep 5:12440

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  37. Marsicano G, Moosmann B, Hermann H, Lutz B, Behl C (2002) Neuroprotective properties of cannabinoids against oxidative stress: role of the cannabinoid receptor CB1. J Neurochem 80:448–456

    CAS  Article  PubMed  Google Scholar 

  38. Martin JB (1999) Molecular basis of the neurodegenerative disorders. N Engl J Med 340:1970–1980

    CAS  Article  PubMed  Google Scholar 

  39. Mattson MP (2004) Metal-catalyzed disruption of membrane protein and lipid signaling in the pathogenesis of neurodegenerative disorders. Ann NY Acad Sci 1012:37–50

    CAS  Article  PubMed  Google Scholar 

  40. Mecha M, Torrao AS, Mestre L, Carrillo-Salinas FJ, Mechoulam R, Guaza C (2012) Cannabidiol protects oligodendrocyte progenitor cells from inflammation-induced apoptosis by attenuating endoplasmic reticulum stress. Cell Death Dis 3:e331

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  41. Micale V, Mazzola C, Drago F (2007) Endocannabinoids and neurodegenerative diseases. Pharm Res 56:382–392

    CAS  Article  Google Scholar 

  42. Milton NG (2002) Anandamide and noladin ether prevent neurotoxicity of the human amyloid-β peptide. Neurosci Lett 332:127–130

    CAS  Article  PubMed  Google Scholar 

  43. Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63

    CAS  Article  PubMed  Google Scholar 

  44. Nakagawa T, Yuan J (2000) Cross-talk between two cysteine protease families. Activation of caspase-12 by calpain in apoptosis. J Cell Biol 150:887–894

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  45. Panikashvili D, Simeonidou C, Bem-Shabat S, Hanus L, Breuer A, Mechoulam R, Shohami E (2001) An endogenous cannabinoid (2-AG) is neuroprotective after brain injury. Nature 413:527–531

    CAS  Article  PubMed  Google Scholar 

  46. Prell T, Lautenschläger J, Weidemann L, Ruhmer J, Witte OW, Grosskreutz J (2014) Endoplasmic reticulum stress is accompanied by activation of NF-κB in amyotrophic lateral sclerosis. J Neuroim 270:29–36

    CAS  Article  Google Scholar 

  47. Ross CA, Poirier MA (2004) Protein aggregation and neurodegenerative disease. Nat Med 10:S10–S17

    Article  PubMed  Google Scholar 

  48. Sagredo O, García-Arencibia M, De Lago E, Finetti S, Decio A, Fernández-Ruiz J (2007) Cannabinoids and neuroprotection in basal ganglia disorders. Mol Neurobiol 36:82–91

    CAS  Article  PubMed  Google Scholar 

  49. Sarker KP, Maruyama I (2003) Anandamide induces cell death independently of cannabinoid receptors or vanilloid receptor 1: possible involvement of lipid rafts. Cell Mol Life Sci 60:1200–1208

    CAS  Article  PubMed  Google Scholar 

  50. Sarne Y, Keren O (2004) Are cannabinoid drugs neurotoxic or neuroprotective? Med Hypotheses 63:187–192

    CAS  Article  PubMed  Google Scholar 

  51. Sarne Y, Mechoulam R (2005) Cannabinoids: between neuroprotection and neurotoxicity. Curr Drug Targets CNS Neurol Disord 4:677–684

    CAS  Article  PubMed  Google Scholar 

  52. Sherman MY, Goldberg AL (2001) Cellular defenses against unfolded proteins: a cell biologist thinks about neurodegenerative diseases. Neuron 29:15–32

    CAS  Article  PubMed  Google Scholar 

  53. Silveira PE, Silveira NA, de Morini VC, Kushmerick C, Naves LA (2010) Opposing effects of cannabinoids and vanilloids on evoked quantal release at the frog neuromuscular junction. Neurosci Lett 473:97–101

    CAS  Article  PubMed  Google Scholar 

  54. Spillantini MG, Schmidt ML, Lee VMY, Trojanowski JQ, Jakes R, Goedert M (1997) α–Synuclein in Lewy bodies. Nature 388:839–840

    CAS  Article  PubMed  Google Scholar 

  55. Stefani IC, Wright D, Polizzi KM, Kontoravdi C (2012) The role of ER stress-induced apoptosis in neurodegeneration. Curr Alzheimer Res 9:373–387

    CAS  Article  PubMed  Google Scholar 

  56. Torrão AS, Café-Mendes CC, Real CC, Hernandes MS, Ferreira AFB, Santos TO, Chaves-Kirsten GP, Mazucanti CHY, Ferro ES, Scavone C, Britto LRG (2012) Different approaches, one target: understanding cellular mechanisms of Parkinson’s and Alzheimer’s diseases. Rev Bras Psiquiatr 34:194–218

    Article  Google Scholar 

  57. Van der Stelt M, Veldhuis WB, Bar PR, Veldink GA, Vliegenthart JF, Nicolay K (2001) Neuroprotection by D9-tetrahydrocannabinol, the main compound in marijuana, against oaubain-induced in vivo excitotoxicity. J Neurosci 21:6475–6579

    Article  PubMed  Google Scholar 

  58. Van der Stelt M, Veldhuis WB, Maccarrone M, Bär PR, Nicolay K, Veldink GA, Di Marzo V, Vliegenthart JFG (2002) Acute neuronal injury, excitotoxicity, and the endocannabinoid system. MolNeurobiol 26:317–346

    Google Scholar 

  59. Van der Stelt M, Di Marzo V (2005) Anandamide as an intracellular messenger regulating ion channel activity. Prostaglandins Other Lipid Mediat 77:111–122

    Article  PubMed  Google Scholar 

  60. Velasco G, Carracedo A, Blázquez C, Lorente M, Aguado T, Haro A, Sánchez C, Galve-Roperh I, Guzmán M (2007) Cannabinoids and gliomas. Mol Neurobiol 36:60–67

    CAS  Article  PubMed  Google Scholar 

  61. Wang X, Fan Z, Wang B, Luo J, Zun-Li K (2007) Activation of double-stranded RNA-activated protein kinase by mild impairment of oxidative metabolism in neurons. J Neurochem 103:2380–2390

    CAS  Article  PubMed  Google Scholar 

  62. Xu C, Bailly-Maitre B, Reed JC (2005) Endoplasmic reticulum stress: cell life and death decisions. J Clin Invest 115:2656–2664

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  63. Yin Y, Sun G, Li E, Kiselyov K, Sun D (2017) ER stress and impaired autophagy flux in neuronal degeneration and brain injury. Ageing Res Rev 34:3–14

    CAS  Article  PubMed  Google Scholar 

  64. Zhang K, Kaufman RJ (2006) The unfolded protein response—a stress signaling pathway critical for health and disease. Neurology 66:S102–S109

    CAS  Article  PubMed  Google Scholar 

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Acknowledgements

This work was supported by the Fundação de Amparo à Pesquisa do Estado de São Paulo-FAPESP (grant number 2014/06372-0). T.A.V. (1279985) and F.C. (1233360) are recipients of Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) fellowships.

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T.A.V. and F.C. designed the experiments, collected and analyzed the data, and wrote the paper. A.P.C. collected, analyzed, and discussed the data. A.S.T. designed the experiments, analyzed and discussed the data, and wrote, edited and commented on the manuscript. All authors approved the final version of the manuscript.

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Correspondence to Andréa S. Torrão.

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The authors declare that they have no conflict of interest.

Financial Support

This work was supported by the Fundação de Amparo à Pesquisa do Estado de São Paulo-FAPESP (grant number 2014/06372-0). T.A.V. (1279985) and F.C. (1233360) are recipients of Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) fellowships.

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Supplementary Fig. 1
figure5

Effects of LPS on ER apoptotic pathway signaling evaluated by immunoblotting method. (a) Effect of ACEA (1.5 μM), AM 251 (4.5 μM), and co-treatment on the PKR phosphorylation in the presence and absence of LPS; ANOVA (F 5,16 = 1.172). (b) Effect of ACEA (1.5 μM), AM 251 (4.5 μM), and co-treatment on eIF2α phosphorylation in the presence and absence of LPS; ANOVA (F 5,30 = 0.58) . (c) Effect of ACEA (1.5 μM), AM 251 (4.5 μM), and co-treatment on the levels of CHOP in the presence and absence of LPS; ANOVA (F 5,27 = 2.392), *p < 0.05 vs.ACEA. (d) Effect of ACEA (1.5 μM), AM251 (4.5 μM), and co-treatment on the levels of Caspase 12 cleavage in the presence and absence of LPS; ANOVA (F 5,25 = 4.279), *p < 0.05,**p < 0.01vs. ACEA.Data (expressed as percentage) represent the mean ± S.E.M. (n = 7–8), analyzed in triplicate and normalized to GAPDH. Statistical significance was determined by ANOVA (one way) followed by a Tukey’s post-hoc test. (GIF 124 kb)

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Vrechi, T.A., Crunfli, F., Costa, A.P. et al. Cannabinoid Receptor Type 1 Agonist ACEA Protects Neurons from Death and Attenuates Endoplasmic Reticulum Stress-Related Apoptotic Pathway Signaling. Neurotox Res 33, 846–855 (2018). https://doi.org/10.1007/s12640-017-9839-1

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Keywords

  • CB1 receptor
  • Cannabinoids
  • Neuroprotection
  • Endoplasmic reticulum
  • Neuroinflammation