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Involvement of PI3K/Akt/FoxO3a and PKA/CREB Signaling Pathways in the Protective Effect of Fluoxetine Against Corticosterone-Induced Cytotoxicity in PC12 Cells

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Abstract

The selective serotonin reuptake inhibitor fluoxetine is neuroprotective in several brain injury models. It is commonly used to treat major depressive disorder and related conditions, but its mechanism of action remains incompletely understood. Activation of the phosphatidylinositol-3-kinase/protein kinase B/forkhead box O3a (PI3K/Akt/FoxO3a) and protein kinase A/cAMP-response element binding protein (PKA/CREB) signaling pathways has been strongly implicated in the pathogenesis of depression and might be the downstream target of fluoxetine. Here, we used PC12 cells exposed to corticosterone (CORT) to study the neuroprotective effects of fluoxetine and the involvement of the PI3K/Akt/FoxO3a and PKA/CREB signaling pathways. Our results show that CORT reduced PC12 cells viability by 70 %, and that fluoxetine showed a concentration-dependent neuroprotective effect. Neuroprotective effects of fluoxetine were abolished by inhibition of PI3K, Akt, and PKA using LY294002, KRX-0401, and H89, respectively. Treatment of PC12 cells with fluoxetine resulted in increased phosphorylation of Akt, FoxO3a, and CREB. Fluoxetine also dose-dependently rescued the phosphorylation levels of Akt, FoxO3a, and CREB, following administration of CORT (from 99 to 110, 56 to 170, 80 to 170 %, respectively). In addition, inhibition of PKA and PI3K/Akt resulted in decreased levels of p-CREB, p-Akt, and p-FoxO3a in the presence of fluoxetine. Furthermore, fluoxetine reversed CORT-induced upregulation of p53-upregulated modulator of apoptosis (Puma) and Bcl-2-interacting mediator of cell death (Bim) via the PI3K/Akt/FoxO3a signaling pathway. H89 treatment reversed the effect of fluoxetine on the mRNA level of brain-derived neurotrophic factor, which was decreased in the presence of CORT. Our data indicate that fluoxetine elicited neuroprotection toward CORT-induced cell death that involves dual regulation from PI3K/Akt/FoxO3a and PKA/CREB pathways.

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Abbreviations

Akt:

Protein kinase B

BDNF:

Brain-derived neurotrophic factor

Bim:

Bcl-2-interacting mediator of cell death

CORT:

Corticosterone

CREB:

cAMP-response element binding protein

DMSO:

Dimethyl sulfoxide

FoxO3a:

Forkhead box O3a

HPA:

Hypothalamic-pituitary-adrenal

MTT:

Methyl thiazolyl tetrazolium

PI3K:

Phosphatidylinositol-3-kinase

PKA:

Protein kinase A

Puma:

p53 upregulated modulator of apoptosis

RPL19:

Ribosomal protein L19

RT-PCR:

Reverse transcription-polymerase chain reaction

References

  • Anacker C, Zunszain PA, Carvalho LA, Pariante CM (2011) The glucocorticoid receptor: pivot of depression and of antidepressant treatment? Psychoneuroendocrinology 36:415–425

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Beaulieu JM (2012) A role for Akt and glycogen synthase kinase-3 as integrators of dopamine and serotonin neurotransmission in mental health. J Psychiatry Neurosci 37:7–16

    Article  PubMed  PubMed Central  Google Scholar 

  • Cecconi D, Mion S, Astner H, Domenici E, Righetti PG, Carboni L (2007) Proteomic analysis of rat cortical neurons after fluoxetine treatment. Brain Res 1135:41–51

    Article  CAS  PubMed  Google Scholar 

  • Chen AC, Shirayama Y, Shin KH, Neve RL, Duman RS (2001) Expression of the cAMP response element binding protein (CREB) in hippocampus produces an antidepressant effect. Biol Psychiatry 49:753–762

    Article  CAS  PubMed  Google Scholar 

  • Chiou SH, Chen SJ, Peng CH, Chang YL, Ku HH, Hsu WM, Ho LL, Lee CH (2006) Fluoxetine up-regulates expression of cellular FLICE-inhibitory protein and inhibits LPS-induced apoptosis in hippocampus-derived neural stem cell. Biochem Biophys Res Commun 343:391–400

    Article  CAS  PubMed  Google Scholar 

  • Ciriaco M, Ventrice P, Russo G, Scicchitano M, Mazzitello G, Scicchitano F, Russo E (2013) Corticosteroid-related central nervous system side effects. J Pharmacol Pharmacother 4:S94–S98

    Article  PubMed  PubMed Central  Google Scholar 

  • Djordjevic A, Djordjevic J, Elakovic I, Adzic M, Matic G, Radojcic MB (2012) Fluoxetine affects hippocampal plasticity, apoptosis and depressive-like behavior of chronically isolated rats. Prog Neuro-Psychopharmacol Biol Psychiatry 36:92–100

    Article  CAS  Google Scholar 

  • Freitas AE, Egea J, Buendia I, Navarro E, Rada P, Cuadrado A, Rodrigues AL, Lopez MG (2015) Agmatine induces Nrf2 and protects against corticosterone effects in hippocampal neuronal cell line. Mol Neurobiol 51:1504–1519

    Article  CAS  PubMed  Google Scholar 

  • Geetha T, Rege SD, Mathews SE, Meakin SO, White MF, Babu JR (2013) Nerve growth factor receptor TrkA, a new receptor in insulin signaling pathway in PC12 cells. J Biol Chem 288:23807–23813

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gu W, Fukuda T, Isaji T, Hashimoto H, Wang Y, Gu J (2013) Alpha1,6-fucosylation regulates neurite formation via the activin/phospho-Smad2 pathway in PC12 cells: the implicated dual effects of Fut8 for TGF-beta/activin-mediated signaling. FASEB J 27:3947–3958

    Article  CAS  PubMed  Google Scholar 

  • Hahn SJ, Choi JS, Rhie DJ, Oh CS, Jo YH, Kim MS (1999) Inhibition by fluoxetine of voltage-activated ion channels in rat PC12 cells. Eur J Pharmacol 367:113–118

    Article  CAS  PubMed  Google Scholar 

  • Ishima T, Fujita Y, Hashimoto K (2014) Interaction of new antidepressants with sigma-1 receptor chaperones and their potentiation of neurite outgrowth in PC12 cells. Eur J Pharmacol 727:167–173

    Article  CAS  PubMed  Google Scholar 

  • Kim HJ, Choi JS, Lee YM, Shim EY, Hong SH, Kim MJ, Min DS, Rhie DJ, Kim MS, Jo YH, Hahn SJ, Yoon SH (2005) Fluoxetine inhibits ATP-induced [Ca(2+)](i) increase in PC12 cells by inhibiting both extracellular Ca(2+) influx and Ca(2+) release from intracellular stores. Neuropharmacology 49:265–274

    Article  CAS  PubMed  Google Scholar 

  • Kim HJ, Kim TH, Choi SJ, Hong YJ, Yang JS, Sung KW, Rhie DJ, Hahn SJ, Yoon SH (2013) Fluoxetine suppresses synaptically induced [Ca(2)(+)]i spikes and excitotoxicity in cultured rat hippocampal neurons. Brain Res 1490:23–34

    Article  CAS  PubMed  Google Scholar 

  • King SC, Tiller AA, Chang AS, Lam DM (1992) Differential regulation of the imipramine-sensitive serotonin transporter by cAMP in human JAr choriocarcinoma cells, rat PC12 pheochromocytoma cells, and C33-14-B1 transgenic mouse fibroblast cells. Biochem Biophys Res Commun 183:487–491

    Article  CAS  PubMed  Google Scholar 

  • Kolla N, Wei Z, Richardson JS, Li XM (2005) Amitriptyline and fluoxetine protect PC12 cells from cell death induced by hydrogen peroxide. J Psychiatry Neurosci 30:196–201

    PubMed  PubMed Central  Google Scholar 

  • Krishnan V, Nestler EJ (2008) The molecular neurobiology of depression. Nature 455:894–902

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lecht S, Arienzakay H, Tabakman R, Jiang H (2007) Dexamethasone-induced down-regulation of nerve growth factor receptor p75NTR is mediated by glucocorticoid type II receptor in PC12 cell model. Open Pharmacol J 19:549–568

    Google Scholar 

  • Lee JY, Kang SR, Yune TY (2015) Fluoxetine prevents oligodendrocyte cell death by inhibiting microglia activation after spinal cord injury. J Neurotrauma 32:633–644

    Article  PubMed  PubMed Central  Google Scholar 

  • Lee JY, Lee HE, Kang SR, Choi HY, Ryu JH, Yune TY (2014) Fluoxetine inhibits transient global ischemia-induced hippocampal neuronal death and memory impairment by preventing blood-brain barrier disruption. Neuropharmacology 79:161–171

    Article  CAS  PubMed  Google Scholar 

  • Li YF, Liu YQ, Huang WC, Luo ZP (2003) Cytoprotective effect is one of common action pathways for antidepressants. Acta Pharmacol Sin 24:996–1000

    CAS  PubMed  Google Scholar 

  • Li YF, Liu YQ, Yang M, Wang HL, Huang WC, Zhao YM, Luo ZP (2004) The cytoprotective effect of inulin-type hexasaccharide extracted from Morinda officinalis on PC12 cells against the lesion induced by corticosterone. Life Sci 75:1531–1538

    Article  CAS  PubMed  Google Scholar 

  • Li YF, Luo ZP (2002) Desipramine antagonized corticosterone-induced apoptosis in cultured PC12 cells. Acta Pharmacol Sin 23:311–314

    CAS  PubMed  Google Scholar 

  • Liang B, Moussaif M, Kuan CJ, Gargus JJ, Sze JY (2006) Serotonin targets the DAF-16/FOXO signaling pathway to modulate stress responses. Cell Metab 4:429–440

    Article  CAS  PubMed  Google Scholar 

  • Ma R, Hu J, Huang C, Wang M, Xiang J, Li G (2014) JAK2/STAT5/Bcl-xL signalling is essential for erythropoietin-mediated protection against apoptosis induced in PC12 cells by the amyloid beta-peptide Abeta25-35. Br J Pharmacol 171:3234–3245

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mao QQ, Huang Z, Ip SP, Xian YF, Che CT (2012) Protective effects of piperine against corticosterone-induced neurotoxicity in PC12 cells. Cell Mol Neurobiol 32:531–537

    Article  CAS  PubMed  Google Scholar 

  • Mendez-David I, Tritschler L, Ali ZE, Damiens MH, Pallardy M, David DJ, Kerdine-Romer S, Gardier AM (2015) Nrf2-signaling and BDNF: a new target for the antidepressant-like activity of chronic fluoxetine treatment in a mouse model of anxiety/depression. Neurosci Lett 597:121–126

    Article  CAS  PubMed  Google Scholar 

  • Morsink MC, Joels M, Sarabdjitsingh RA, Meijer OC, De Kloet ER, Datson NA (2006) The dynamic pattern of glucocorticoid receptor-mediated transcriptional responses in neuronal PC12 cells. J Neurochem 99:1282–1298

    Article  CAS  PubMed  Google Scholar 

  • Pittenger C, Duman RS (2008) Stress, depression, and neuroplasticity: a convergence of mechanisms. Neuropsychopharmacology 33:88–109

    Article  CAS  PubMed  Google Scholar 

  • Plattner F, Hayashi K, Hernandez A, Benavides DR, Tassin TC, Tan C, Day J, Fina MW, Yuen EY, Yan Z, Goldberg MS, Nairn AC, Greengard P, Nestler EJ, Taussig R, Nishi A, Houslay MD, Bibb JA (2015) The role of ventral striatal cAMP signaling in stress-induced behaviors. Nat Neurosci 18:1094–1100

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Polman JA, Welten JE, Bosch DS, de Jonge RT, Balog J, van der Maarel SM, de Kloet ER, Datson NA (2012) A genome-wide signature of glucocorticoid receptor binding in neuronal PC12 cells. BMC Neurosci 13:118

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Polter A, Yang S, Zmijewska AA, van Groen T, Paik JH, Depinho RA, Peng SL, Jope RS, Li X (2009) Forkhead box, class O transcription factors in brain: regulation and behavioral manifestation. Biol Psychiatry 65:150–159

    Article  CAS  PubMed  Google Scholar 

  • Qi X, Lin W, Li J, Li H, Wang W, Wang D, Sun M (2008) Fluoxetine increases the activity of the ERK-CREB signal system and alleviates the depressive-like behavior in rats exposed to chronic forced swim stress. Neurobiol Dis 31:278–285

    Article  CAS  PubMed  Google Scholar 

  • Sanphui P, Biswas SC (2013) FoxO3a is activated and executes neuron death via Bim in response to beta-amyloid. Cell Death Dis 4:e625

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sapolsky RM, Krey LC, McEwen BS (1984) Glucocorticoid-sensitive hippocampal neurons are involved in terminating the adrenocortical stress response. Proc Natl Acad Sci U S A 81:6174–6177

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Shin LM, Liberzon I (2010) The neurocircuitry of fear, stress, and anxiety disorders. Neuropsychopharmacology 35:169–191

    Article  PubMed  Google Scholar 

  • Subramaniam S, Shahani N, Strelau J, Laliberte C, Brandt R, Kaplan D, Unsicker K (2005) Insulin-like growth factor 1 inhibits extracellular signal-regulated kinase to promote neuronal survival via the phosphatidylinositol 3-kinase/protein kinase A/c-Raf pathway. J Neurosci 25:2838–2852

    Article  CAS  PubMed  Google Scholar 

  • Suzuki A, Fukushima H, Mukawa T, Toyoda H, Wu LJ, Zhao MG, Xu H, Shang Y, Endoh K, Iwamoto T, Mamiya N, Okano E, Hasegawa S, Mercaldo V, Zhang Y, Maeda R, Ohta M, Josselyn SA, Zhuo M, Kida S (2011) Upregulation of CREB-mediated transcription enhances both short- and long-term memory. J Neurosci 31:8786–8802

    Article  CAS  PubMed  Google Scholar 

  • Tillinger A, Nostramo R, Kvetnansky R, Serova L, Sabban EL (2013) Stress-induced changes in gene expression of urocortin 2 and other CRH peptides in rat adrenal medulla: involvement of glucocorticoids. J Neurochem 125:185–192

    Article  CAS  PubMed  Google Scholar 

  • Tiraboschi E, Tardito D, Kasahara J, Moraschi S, Pruneri P, Gennarelli M, Racagni G, Popoli M (2004) Selective phosphorylation of nuclear CREB by fluoxetine is linked to activation of CaM kinase IV and MAP kinase cascades. Neuropsychopharmacology 29:1831–1840

    Article  CAS  PubMed  Google Scholar 

  • Wakamatsu Y, Zhao X, Jin C, Day N, Shibahara M, Nomura N, Nakahara T, Murata T, Yokoyama KK (2001) Mannosylerythritol lipid induces characteristics of neuronal differentiation in PC12 cells through an ERK-related signal cascade. Eur J Biochem 268:374–383

    Article  CAS  PubMed  Google Scholar 

  • Wang J, Sun P, Bao Y, Liu J, An L (2011) Cytotoxicity of single-walled carbon nanotubes on PC12 cells. Toxicol in Vitro 25:242–250

    Article  CAS  PubMed  Google Scholar 

  • Wang H, Duan X, Ren Y, Liu Y, Huang M, Liu P, Wang R, Gao G, Zhou L, Feng Z, Zheng W (2013a) FoxO3a negatively regulates nerve growth factor-induced neuronal differentiation through inhibiting the expression of neurochondrin in PC12 cells. Mol Neurobiol 47:24–36

    Article  CAS  PubMed  Google Scholar 

  • Wang H, Zhou X, Huang J, Mu N, Guo Z, Wen Q, Wang R, Chen S, Feng ZP, Zheng W (2013b) The role of Akt/FoxO3a in the protective effect of venlafaxine against corticosterone-induced cell death in PC12 cells. Psychopharmacology 228:129–141

    Article  CAS  PubMed  Google Scholar 

  • Wang H, Liao S, Geng R, Zheng Y, Liao R, Yan F, Thrimawithana T, Little PJ, Feng ZP, Lazarovici P, Zheng W (2015a) IGF-1 signaling via the PI3K/Akt pathway confers neuroprotection in human retinal pigment epithelial cells exposed to sodium nitroprusside insult. J Mol Neurosci 55:931–940

    Article  CAS  PubMed  Google Scholar 

  • Wang H, Quirion R, Little PJ, Cheng Y, Feng ZP, Sun HS, Xu J, Zheng W (2015b) Forkhead box O transcription factors as possible mediators in the development of major depression. Neuropharmacology 99:527–537

    Article  CAS  PubMed  Google Scholar 

  • Zheng WH, Kar S, Quirion R (2002) Insulin-like growth factor-1-induced phosphorylation of transcription factor FKHRL1 is mediated by phosphatidylinositol 3-kinase/Akt kinase and role of this pathway in insulin-like growth factor-1-induced survival of cultured hippocampal neurons. Mol Pharmacol 62:225–233

    Article  CAS  PubMed  Google Scholar 

  • Zheng WH, Quirion R (2006) Insulin-like growth factor-1 (IGF-1) induces the activation/phosphorylation of Akt kinase and cAMP response element-binding protein (CREB) by activating different signaling pathways in PC12 cells. BMC Neurosci 7:51

    Article  PubMed  PubMed Central  Google Scholar 

  • Zhou H, Li X, Gao M (2009) Curcumin protects PC12 cells from corticosterone-induced cytotoxicity: possible involvement of the ERK1/2 pathway. Basic Clin Pharmacol Toxicol 104:236–240

    Article  PubMed  Google Scholar 

  • Zhou LJ, Zhu XZ (2000) Reactive oxygen species-induced apoptosis in PC12 cells and protective effect of bilobalide. J Pharmacol Exp Ther 293:982–988

    CAS  PubMed  Google Scholar 

  • Zhou W, Chen L, Yang S, Li F, Li X (2012) Behavioral stress-induced activation of FoxO3a in the cerebral cortex of mice. Biol Psychiatry 71:583–592

    Article  CAS  PubMed  Google Scholar 

  • Zhu MY, Ordway GA (1997) Down-regulation of norepinephrine transporters on PC12 cells by transporter inhibitors. J Neurochem 68:134–141

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This research was supported by National Natural Science Foundation of China (Nos. 81301099 and 81373384), Natural Science Foundation of Guangdong Province (No. S2013040014202), China Postdoctoral Science Foundation (No. 2013M542192), and National Science and Technology Major Projects for “Major New Drugs Innovation and Development” (No. 2012ZX09J1211003C).

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Authors and Affiliations

  1. Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, 510515, China

    Bingqing Zeng, Yiwen Li, Bo Niu, Xinyi Wang, Yufang Cheng, Zhongzhen Zhou, Tingting You, Haitao Wang & Jiangping Xu

  2. Department of Pharmacy, Guangdong Provincial Corps Hospital of Chinese People’s Armed Police Forces, Guangzhou, 510507, China

    Yonggang Liu

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  1. Bingqing Zeng
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  2. Yiwen Li
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Correspondence to Haitao Wang or Jiangping Xu.

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Zeng, B., Li, Y., Niu, B. et al. Involvement of PI3K/Akt/FoxO3a and PKA/CREB Signaling Pathways in the Protective Effect of Fluoxetine Against Corticosterone-Induced Cytotoxicity in PC12 Cells. J Mol Neurosci 59, 567–578 (2016). https://doi.org/10.1007/s12031-016-0779-7

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