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Duloxetine Protects Human Neuroblastoma Cells from Oxidative Stress-Induced Cell Death Through Akt/Nrf-2/HO-1 Pathway

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Abstract

The contribution of oxidative stress to the pathophysiology of depression has been described in numerous studies. Particularly, an increased production of reactive oxygen species (ROS) caused by mitochondrial dysfunction can lead to neuronal cell death. Human neuroblastoma SH-SY5Y cells were used to investigate the neuroprotective effect of the antidepressant duloxetine against rotenone-induced oxidative stress. SH-SY5Y cells were pretreated with duloxetine (1–5 µM) for 24 h followed by a 24-h rotenone exposure (10 µM). The phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) inhibitor LY294002 (10 µM) and the heme oxygenase 1 (HO-1) inhibitor zinc protoporphyrin IX-ZnPP (5 µM) were added to cultures 1 h prior duloxetine treatments. After treatments cell viability and ROS generation were assessed. NF-E2-related factor-2 (Nrf2) nuclear translocation was assessed by immunofluorescent staining after 4 and 8 h of duloxetine incubation. Furthermore, the Nrf2 and HO-1 mRNA expression was carried out after 4–48 h of duloxetine treatment by qRT-PCR. Duloxetine pretreatment antagonized rotenone-induced overproduction of ROS and cell death in SH-SY5Y cells. In addition, a 1-h pretreatment with LY294002 abolished duloxetine’s protective effect. Duloxetine also induced nuclear translocation of the Nrf2 and the expression of its target gene, HO-1. Finally, the HO-1 inhibitor, ZnPP, suppressed the duloxetine protective effect. Overall, these results indicate that the mechanism of duloxetine neuroprotective action against oxidative stress and cell death might rely on the Akt/Nrf2/HO-1 pathways.

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Acknowledgements

This study was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), FAPESC (TO 22.333/2010-6). Rede IBN-Net/CNPq (IBN 01.06.0842-00), CAPES-PROCAD, CAPES-REUNI, Coordenação de Aperfeiçoamento de Pessoal de Nível Superior-CAPES. INCT-National Institute of Science and Technology for Excitotoxicity and Neuroprotection.

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  1. Departamento de Bioquímica, CCB, Universidade Federal de Santa Catarina, Campus Universitário, Florianópolis, SC, 88040-900, Brazil

    Daiane Fátima Engel, Vicente Lieberknecht, Ana Lucia Severo Rodrigues, Andreza Fabro de Bem & Nelson Horácio Gabilan

  2. Universidade do Extremo Sul Catarinense, Criciúma, SC, Brazil

    Jade de Oliveira

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  1. Daiane Fátima Engel
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Correspondence to Daiane Fátima Engel.

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Engel, D.F., de Oliveira, J., Lieberknecht, V. et al. Duloxetine Protects Human Neuroblastoma Cells from Oxidative Stress-Induced Cell Death Through Akt/Nrf-2/HO-1 Pathway. Neurochem Res 43, 387–396 (2018). https://doi.org/10.1007/s11064-017-2433-3

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