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

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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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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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  1. Department of Physiology and Biophysics, Institute of Biomedical Sciences, University of São Paulo, Av. Prof. Lineu Prestes, 1524, São Paulo, SP, 05508-000, Brazil

    Talita A. Vrechi, Fernanda Crunfli, Andressa P. Costa & Andréa S. Torrão

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  1. Talita A. Vrechi
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  2. Fernanda Crunfli
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  3. Andressa P. Costa
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  4. Andréa S. Torrão
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Contributions

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

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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