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Agomelatine and Duloxetine Synergistically Modulates Apoptotic Pathway by Inhibiting Oxidative Stress Triggered Intracellular Calcium Entry in Neuronal PC12 Cells: Role of TRPM2 and Voltage-Gated Calcium Channels

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Abstract

Calcium ion (Ca2+) is one of the universal second messengers, which acts in a wide range of cellular processes. Results of recent studies indicated that ROS generated by depression leads to loss of endoplasmic reticulum-Ca2+ homeostasis, oxidative stress, and apoptosis. Agomelatine and duloxetine are novel antidepressant and antioxidant drugs and may reduce oxidative stress, apoptosis, and Ca2+ entry through TRPM2 and voltage-gated calcium channels. We tested the effects of agomelatine, duloxetine, and their combination on oxidative stress, Ca2+ influx, mitochondrial depolarization, apoptosis, and caspase values in the PC-12 neuronal cells. PC-12 neuronal cells were exposed in cell culture and exposed to appropriate non-toxic concentrations and incubation times for agomelatine were determined in the neurons by assessing cell viability. Then PC-12 cells were incubated with agomelatine and duloxetine for 24 h. Treatment of cultured PC-12 cells with agomelatine, duloxetine, and their combination results in a protection on apoptosis, caspase-3, caspase-9, mitochondrial membrane depolarization, cytosolic ROS production, glutathione peroxidase, reduced glutathione, and lipid peroxidation, values. Ca2+ entry through non-specific TRPM2 channel blocker (2-APB) and voltage-gated Ca2+ channel blockers (verapamil and diltiazem) was modulated by agomelatine and duloxetine. However, effects of duloxetine on the Ca2+ entry through TRPM2 channels were higher than in agomelatine. Results of current study suggest that the agomelatine and duloxetine are useful against apoptotic cell death and oxidative stress in PC-12 cells, which seem to be dependent on mitochondrial damage and increased levels of intracellular Ca2+ through activation of TRPM2 and voltage-gated Ca2+ channels.

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Abbreviations

[Ca2+]i :

Intracellular Ca2+

2-APB:

2-Aminoethyl diphenylborinate

CNS:

Central nervous system

DMSO:

Dimethyl sulfoxide

DTNB:

5,5′-Dithiobis-(2-nitrobenzoic acid)

GSH:

Glutathione

GSH-Px:

Glutathione peroxidase

LP:

Lipid peroxidation

MDD:

Major depressive disorder

NMDG+ :

N-Methyl-d-glucamine

ROS:

Reactive oxygen species

TRP:

Transient receptor potential

TRPM2:

Transient receptor potential melastatin

VGCC:

Voltage-gated calcium channel

H2O2 :

Hydrogen peroxide

References

  • Aguiar CC, Almeida AB, Araujo PV, Vasconcelos GS, Chaves EM, do Vale OC, Macedo DS, Leal LK, de Barros Viana GS, Vasconcelos SM (2013) Effects of agomelatine on oxidative stress in the brain of mice after chemically induced seizures. Cell Mol Neurobiol 33:825–835

    Article  PubMed  Google Scholar 

  • Altinkilic S, Naziroglu M, Uguz AC, Ozcankaya R (2010) Fish oil and antipsychotic drug risperidone modulate oxidative stress in PC12 cell membranes through regulation of cytosolic calcium ion release and antioxidant system. J Membr Biol 235:211–218

    Article  CAS  PubMed  Google Scholar 

  • Amaral MD, Pozzo-Miller L (2007) BDNF induces calcium elevations associated with IBDNF, a nonselective cationic current mediated by TRPC channels. J Neurophysiol 98:2476–2482

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Bains JS, Shaw CA (1997) Neurodegenerative disorders in humans: the role of glutathione in oxidative stress-mediated neuronal death. Brain Res Brain Res Rev 25:335–358

    Article  CAS  PubMed  Google Scholar 

  • Beales IL, Ogunwobi OO (2010) Microsomal prostaglandin E synthase-1 inhibition blocks proliferation and enhances apoptosis in oesophageal adenocarcinoma cells without affecting endothelial prostacyclin production. Int J Cancer 126:2247–2255

    CAS  PubMed  Google Scholar 

  • Berliocchi L, Bano D, Nicotera P (2005) Ca2+ signals and death programmes in neurons. Philos Trans R Soc Lond B Biol Sci 360:2255–2258

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Birnbaumer L, Zhu X, Jiang M, Boulay G, Peyton M, Vannier B, Brown D, Platano D, Sadeghi H, Stefani E, Birnbaumer M (1996) On the molecular basis and regulation of cellular capacitative calcium entry: roles for Trp proteins. Proc Natl Acad Sci USA 93:15195–15202

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Bramham CR, Messaoudi E (2005) BDNF function in adult synaptic plasticity: the synaptic consolidation hypothesis. Prog Neurobiol 76:99–125

    Article  CAS  PubMed  Google Scholar 

  • Carter NJ, McCormack PL (2009) Duloxetine: a review of its use in the treatment of generalized anxiety disorder. CNS Drugs 23:523–541

    Article  CAS  PubMed  Google Scholar 

  • Castedo M, Hirsch T, Susin SA, Zamzami N, Marchetti P, Macho A, Kroemer G (1996) Sequential acquisition of mitochondrial and plasma membrane alterations during early lymphocyte apoptosis. J Immunol 157:512–521

    CAS  PubMed  Google Scholar 

  • Clapham DE (2003) TRP channels as cellular sensors. Nature 426:517–524

    Article  CAS  PubMed  Google Scholar 

  • De Berardis D, Fornaro M, Serroni N, Olivieri L, Marini S, Moschetta FS, Srinivasan V, Assetta M, Valchera A, Salone A, Martinotti G, Onofrj M, Di Giannantonio M (2013) Agomelatine treatment of major depressive disorder in Parkinson’s disease: a case series. J Neuropsychiatry Clin Neurosci 25:343–345

    Article  PubMed  Google Scholar 

  • Duman RS, Malberg J, Thome J (1999) Neural plasticity to stress and antidepressant treatment. Biol Psychiatry 46:1181–1191

    Article  CAS  PubMed  Google Scholar 

  • Eren I, Naziroglu M, Demirdas A, Celik O, Uguz AC, Altunbasak A, Ozmen I, Uz E (2007) Venlafaxine modulates depression-induced oxidative stress in brain and medulla of rat. Neurochem Res 32:497–505

    Article  CAS  PubMed  Google Scholar 

  • Finkbeiner S, Tavazoie SF, Maloratsky A, Jacobs KM, Harris KM, Greenberg ME (1997) CREB: a major mediator of neuronal neurotrophin responses. Neuron 19:1031–1047

    Article  CAS  PubMed  Google Scholar 

  • Fonfria E, Marshall IC, Boyfield I, Skaper SD, Hughes JP, Owen DE, Zhang W, Miller BA, Benham CD, McNulty S (2005) Amyloid beta-peptide(1-42) and hydrogen peroxide-induced toxicity are mediated by TRPM2 in rat primary striatal cultures. J Neurochem 95:715–723

    Article  CAS  PubMed  Google Scholar 

  • Gao Y, Lei Z, Lu C, Roisen FJ, El-Mallakh RS (2010) Effect of ionic stress on apoptosis and the expression of TRPM2 in human olfactory neuroepithelial-derived progenitors. World J Biol Psychiatry 11:972–984

    Article  PubMed  Google Scholar 

  • Grynkiewicz G, Poenie M, Tsien RY (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 260:3440–3450

    CAS  PubMed  Google Scholar 

  • Herken H, Akyol O, Yilmaz HR, Tutkun H, Savas HA, Ozen ME, Kalenderoglu A, Gulec M (2006) Nitric oxide, adenosine deaminase, xanthine oxidase and superoxide dismutase in patients with panic disorder: alterations by antidepressant treatment. Hum Psychopharmacol 21:53–59

    Article  CAS  PubMed  Google Scholar 

  • Hovatta I, Juhila J, Donner J (2010) Oxidative stress in anxiety and comorbid disorders. Neurosci Res 68:261–275

    Article  CAS  PubMed  Google Scholar 

  • Korkmaz S, Erturan I, Naziroglu M, Uguz AC, Cig B, Ovey IS (2011) Colchicine modulates oxidative stress in serum and neutrophil of patients with Behcet disease through regulation of Ca(2) release and antioxidant system. J Membr Biol 244:113–120

    Article  CAS  PubMed  Google Scholar 

  • Lawrence RA, Burk RF (1976) Glutathione peroxidase activity in selenium-deficient rat liver. Biochem Biophys Res Commun 71:952–958

    Article  CAS  PubMed  Google Scholar 

  • Li YX, Zhang Y, Lester HA, Schuman EM, Davidson N (1998) Enhancement of neurotransmitter release induced by brain-derived neurotrophic factor in cultured hippocampal neurons. J Neurosci 18:10231–10240

    CAS  PubMed  Google Scholar 

  • Li GY, Fan B, Zheng YC (2010) Calcium overload is a critical step in programmed necrosis of ARPE-19 cells induced by high-concentration HO. Biomed Environ Sci 23:371–377

    Article  CAS  PubMed  Google Scholar 

  • Liu HC, Chen GG, Vlantis AC, Tse GM, Chan AT, van Hasselt CA (2008) Inhibition of apoptosis in human laryngeal cancer cells by E6 and E7 oncoproteins of human papillomavirus 16. J Cell Biochem 103:1125–1143

    Article  CAS  PubMed  Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    CAS  PubMed  Google Scholar 

  • Maes M, De Vos N, Pioli R, Demedts P, Wauters A, Neels H, Christophe A (2000) Lower serum vitamin E concentrations in major depression. Another marker of lowered antioxidant defenses in that illness. J Affect Disord 58:241–246

    Article  CAS  PubMed  Google Scholar 

  • Manji HK, Gottesman II, Gould TD (2003) Signal transduction and genes-to-behaviors pathways in psychiatric diseases. Sci STKE 2003:pe49

  • Michel TM, Pulschen D, Thome J (2012) The role of oxidative stress in depressive disorders. Curr Pharm Des 18:5890–5899

    Article  CAS  PubMed  Google Scholar 

  • Mikoshiba K (1997) The InsP3 receptor and intracellular Ca2+ signaling. Curr Opin Neurobiol 7:339–345

    Article  CAS  PubMed  Google Scholar 

  • Naziroglu M (2007) New molecular mechanisms on the activation of TRPM2 channels by oxidative stress and ADP-ribose. Neurochem Res 32:1990–2001

    Article  CAS  PubMed  Google Scholar 

  • Naziroglu M (2009) Role of selenium on calcium signaling and oxidative stress-induced molecular pathways in epilepsy. Neurochem Res 34:2181–2191

    Article  CAS  PubMed  Google Scholar 

  • Naziroglu M, Luckhoff A (2008a) A calcium influx pathway regulated separately by oxidative stress and ADP-ribose in TRPM2 channels: single channel events. Neurochem Res 33:1256–1262

    Article  CAS  PubMed  Google Scholar 

  • Naziroglu M, Luckhoff A (2008b) Effects of antioxidants on calcium influx through TRPM2 channels in transfected cells activated by hydrogen peroxide. J Neurol Sci 270:152–158

    Article  CAS  PubMed  Google Scholar 

  • Naziroglu M, Cig B, Ozgul C (2013) Neuroprotection induced by N-acetylcysteine against cytosolic glutathione depletion-induced Ca2+ influx in dorsal root ganglion neurons of mice: role of TRPV1 channels. Neuroscience 242:151–160

    Article  CAS  PubMed  Google Scholar 

  • Ozgul C, Naziroglu M (2012) TRPM2 channel protective properties of N-acetylcysteine on cytosolic glutathione depletion dependent oxidative stress and Ca2+ influx in rat dorsal root ganglion. Physiol Behav 106:122–128

    Article  CAS  PubMed  Google Scholar 

  • Pang DB, Hong J (2010) [Ca(2+)]i homeostasis and caspase-3 gene expression in verapamil-induced retinal pigment epithelium cells apoptosis in vitro. Int J Ophthalmol 3:288–290

    PubMed Central  PubMed  Google Scholar 

  • Placer ZA, Cushman LL, Johnson BC (1966) Estimation of product of lipid peroxidation (malonyl dialdehyde) in biochemical systems. Anal Biochem 16:359–364

    Article  CAS  PubMed  Google Scholar 

  • Prickaerts J, De Vry J, Boere J, Kenis G, Quinton MS, Engel S, Melnick L, Schreiber R (2012) Differential BDNF responses of triple versus dual reuptake inhibition in neuronal and astrocytoma cells as well as in rat hippocampus and prefrontal cortex. J Mol Neurosci 48:167–175

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Putney JW Jr (2003) Capacitative calcium entry in the nervous system. Cell Calcium 34:339–344

    Article  CAS  PubMed  Google Scholar 

  • Qi H, Chen HZ, Jin ZJ (2002) Caspase 3 gene expression and [Ca2+]i homeostasis underlying desipramine-induced C6 glioma cell apoptosis. Acta Pharmacol Sin 23:803–807

    CAS  PubMed  Google Scholar 

  • Rothe G, Oser A, Valet G (1988) Dihydrorhodamine 123: a new flow cytometric indicator for respiratory burst activity in neutrophil granulocytes. Naturwissenschaften 75:354–355

    Article  CAS  PubMed  Google Scholar 

  • Soumier A, Banasr M, Lortet S, Masmejean F, Bernard N, Kerkerian-Le-Goff L, Gabriel C, Millan MJ, Mocaer E, Daszuta A (2009) Mechanisms contributing to the phase-dependent regulation of neurogenesis by the novel antidepressant, agomelatine, in the adult rat hippocampus. Neuropsychopharmacology 34:2390–2403

    Article  CAS  PubMed  Google Scholar 

  • Srinivasan V, Zakaria R, Othman Z, Lauterbach EC, Acuna-Castroviejo D (2012) Agomelatine in depressive disorders: its novel mechanisms of action. J Neuropsychiatry Clin Neurosci 24:290–308

    Article  CAS  PubMed  Google Scholar 

  • Staaf S, Franck MC, Marmigere F, Mattsson JP, Ernfors P (2010) Dynamic expression of the TRPM subgroup of ion channels in developing mouse sensory neurons. Gene Expr Patterns 10:65–74

    Article  CAS  PubMed  Google Scholar 

  • Su HC, Ma CT, Yu BC, Chien YC, Tsai CC, Huang WC, Lin CF, Chuang YH, Young KC, Wang JN, Tsao CW (2012) Glycogen synthase kinase-3beta regulates anti-inflammatory property of fluoxetine. Int Immunopharmacol 14:150–156

    Article  CAS  PubMed  Google Scholar 

  • Uguz AC, Naziroglu M (2012) Effects of selenium on calcium signaling and apoptosis in rat dorsal root ganglion neurons induced by oxidative stress. Neurochem Res 37:1631–1638

    Article  CAS  PubMed  Google Scholar 

  • Uguz AC, Naziroglu M, Espino J, Bejarano I, Gonzalez D, Rodriguez AB, Pariente JA (2009) Selenium modulates oxidative stress-induced cell apoptosis in human myeloid HL-60 cells through regulation of calcium release and caspase-3 and -9 activities. J Membr Biol 232:15–23

    Article  CAS  PubMed  Google Scholar 

  • Uguz AC, Cig B, Espino J, Bejarano I, Naziroglu M, Rodriguez AB, Pariente JA (2012) Melatonin potentiates chemotherapy-induced cytotoxicity and apoptosis in rat pancreatic tumor cells. J Pineal Res 53:91–98

    Article  CAS  PubMed  Google Scholar 

  • Wang H, Yuan G, Prabhakar NR, Boswell M, Katz DM (2006) Secretion of brain-derived neurotrophic factor from PC12 cells in response to oxidative stress requires autocrine dopamine signaling. J Neurochem 96:694–705

    Article  CAS  PubMed  Google Scholar 

  • Warner-Schmidt JL, Duman RS (2007) VEGF is an essential mediator of the neurogenic and behavioral actions of antidepressants. Proc Natl Acad Sci USA 104:4647–4652

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Xie YF, Belrose JC, Lei G, Tymianski M, Mori Y, Macdonald JF, Jackson MF (2011) Dependence of NMDA/GSK-3beta mediated metaplasticity on TRPM2 channels at hippocampal CA3-CA1 synapses. Mol Brain 4:44

    Article  PubMed Central  CAS  PubMed  Google Scholar 

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Acknowledgments

There is no financial support for the current study. Special thanks to Stephanie Mayo and Eleni Mae Lazarou for polishing the English.

Conflict of interest

None of the authors has any conflicts to disclose. All authors approved the final manuscript.

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Authors and Affiliations

  1. Department of Psychiatry, Faculty of Medicine, University of Suleyman Demirel, Isparta, Turkey

    Abdullah Akpinar

  2. Department of Biophysics, Faculty of Medicine, University of Suleyman Demirel, Isparta, Turkey

    Abdülhadi Cihangir Uğuz & Mustafa Nazıroğlu

  3. Neuroscience Research Center, University of Suleyman Demirel, Isparta, Turkey

    Abdülhadi Cihangir Uğuz & Mustafa Nazıroğlu

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  1. Abdullah Akpinar
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  2. Abdülhadi Cihangir Uğuz
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  3. Mustafa Nazıroğlu
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Correspondence to Abdülhadi Cihangir Uğuz.

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Akpinar, A., Uğuz, A.C. & Nazıroğlu, M. Agomelatine and Duloxetine Synergistically Modulates Apoptotic Pathway by Inhibiting Oxidative Stress Triggered Intracellular Calcium Entry in Neuronal PC12 Cells: Role of TRPM2 and Voltage-Gated Calcium Channels. J Membrane Biol 247, 451–459 (2014). https://doi.org/10.1007/s00232-014-9652-1

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