Neurochemical Research

, Volume 43, Issue 1, pp 153–161 | Cite as

Comparing the Effects of Melatonin with Caloric Restriction in the Hippocampus of Aging Mice: Involvement of Sirtuin1 and the FOXOs Pathway

  • Anorut Jenwitheesuk
  • Seongjoon Park
  • Prapimpun Wongchitrat
  • Jiraporn Tocharus
  • Sujira Mukda
  • Isao ShimokawaEmail author
  • Piyarat GovitrapongEmail author
Original Paper


It has been suggested that age-related neurodegeneration might be associated with neuropeptide Y (NPY); sirtuin1 (SIRT1) and forkhead box transcription factors O subfamily (FOXOs) pathways. Melatonin, a hormone mainly secreted by the pineal gland, is another anti-aging agent associated with the SIRT1-FOXOs pathway. This study aimed to compare the effects of melatonin (Mel) and caloric restriction (CR) on the expression of Sirt1, FoxO1, FoxO3a and FOXOs target genes in the aging mouse hippocampus. Neuropeptide Y-knockout (NpyKO) and wild-type (WT) male mice aged 19 months were previously treated either with food ad libitum or CR for 16 months. WT old animals were divided into four groups: control, CR, Mel and CR+Mel treated groups. The Mel and CR+Mel were treated with melatonin 10 mg/kg, daily, subcutaneously for 7 consecutive days. Mel treatment upregulated the mRNA expression of Sirt1, FOXOs (FoxO1 and FoxO3a) target genes that regulated the cell cycle [e.g., cyclin-dependent kinase inhibitor 1B (p27)], Wingless and INT-1 (Wnt1) and inducible signaling pathway protein 1 (Wisp1) in the aged mouse hippocampus. CR treatment also showed the similar actions. However, the mRNA expression of Sirt1, FoxO1, FoxO3a, p27 or Wisp1 did not alter in the CR+Mel group when compared with CR or Mel group. Melatonin could not produce any additive effect on the CR treatment group, suggesting that both treatments mimicked the effect, possibly via the same pathway. NPY which mediates physiological adaptations to energy deficits is an essential link between CR and longevity in mice. In order to focus on the role of Npy in mediating the effects of melatonin, the gene expression between NpyKO and WT male mice were compared. Our data showed that, in the absence of Npy, melatonin could not mediate effects on those gene expressions, suggesting that Npy was required for melatonin to mediate the effect, possibly, on life extension.


Caloric restriction Melatonin Sirtuin1 FoxO1 Neuropeptide Y Aging 



A disintegrin and metalloprotease domain 10


Agouti-related peptide


Adenosine monophosphate-activated protein kinase


Cocaine and amphetamine regulated transcript


CREB-binding protein


Caloric restriction


Wnt inhibitor Dickkopf WNT signaling pathway inhibitor 1


Forkhead box transcription factors and the O subfamily


Glyceraldehyde 3-phosphate dehydrogenase


Insulin-like growth factor-1




Nicotinamide adenine dinucleotide


Nicotinamide phosphoribosyltransferase


Nuclear factor-κB


Nicotinamide/nicotinic acid mononucleotide adenylyltransferase


Neuropeptide Y


Nuclear factor (erythroid-derived 2)-like 2


Anorexigenic proopiomelanocortin


Cyclin-dependent kinase inhibitor 1B


E1A binding protein p300


Tumor suppressor protein 53


Peroxisome proliferator-activated receptor gamma coactivator 1-alpha






Peroxisome proliferator-activated receptor gamma


Senescence-accelerated mouse prone 8


Senescence-accelerated-resistant mouse




Wn1 inducible pathway protein1





This study was supported in part by the Thailand Research Fund (TRF) (DPG5780001), a Mahidol University Research Grant to PG and Goho Life Sciences International Funding to AJ.


  1. 1.
    Morris JK, Bomhoff GL, Stanford JA, Geiger PC (2010) Neurodegeneration in an animal model of Parkinson’s disease is exacerbated by a high-fat diet. Am J Physiol Regul Integr Comp Physiol 299:R1082–R1090Google Scholar
  2. 2.
    Rotermund C, Truckenmuller FM, Schell H, Kahle PJ (2014) Diet-induced obesity accelerates the onset of terminal phenotypes in alpha-synuclein transgenic mice. J Neurochem 131:848-858Google Scholar
  3. 3.
    le Coutre J, Mattson MP, Dillin A, Friedman J, Bistrian B (2013) Nutrition and the biology of human ageing: cognitive decline/food intake and caloric restriction. J Nutr Health Aging 17:717–720Google Scholar
  4. 4.
    Wu P, Shen Q, Dong S, Xu Z, Tsien JZ, Hu Y (2008) Calorie restriction ameliorates neurodegenerative phenotypes in forebrain-specific presenilin-1 and presenilin-2 double knockout mice. Neurobiol Aging 29:1502–1511Google Scholar
  5. 5.
    Van Cauwenberghe C, Vandendriessche C, Libert C, Vandenbroucke RE (2016) Caloric restriction: beneficial effects on brain aging and Alzheimer’s disease. Mamm Genome 27:300–319Google Scholar
  6. 6.
    Maiese K, Chong ZZ, Shang YC, Wang S (2011) Translating cell survival and cell longevity into treatment strategies with SIRT1. Rom J Morphol Embryol 52:1173–1185Google Scholar
  7. 7.
    Narasimhan SD, Yen K, Tissenbaum HA (2009) Converging pathways in lifespan regulation. Curr Biol 19:R657-R666Google Scholar
  8. 8.
    Quintas A, de Solis AJ, Diez-Guerra FJ, Carrascosa JM, Bogonez E (2012) Age-associated decrease of SIRT1 expression in rat hippocampus: prevention by late onset caloric restriction. Exp Gerontol 47:198–201Google Scholar
  9. 9.
    Furuyama T, Yamashita H, Kitayama K, Higami Y, Shimokawa I, Mori N (2002) Effects of aging and caloric restriction on the gene expression of Foxo1, 3, and 4 (FKHR, FKHRL1, and AFX) in the rat skeletal muscles. Microsc Res Tech 59:331–334Google Scholar
  10. 10.
    Yamaza H, Chiba T, Higami Y, Shimokawa I (2002) Lifespan extension by caloric restriction: an aspect of energy metabolism. Microsc Res Tech 59:325–330Google Scholar
  11. 11.
    Fukunaga K, Shioda N (2009) Pathophysiological relevance of forkhead transcription factors in brain ischemia. Adv Exp Med Biol 665:130–142Google Scholar
  12. 12.
    Zemva J, Schilbach K, Stohr O, Moll L, Franko A, Krone W, Wiesner RJ, Schubert M (2012) Central FoxO3a and FoxO6 expression is down-regulated in obesity induced diabetes but not in aging. Exp Clin Endocrinol Diabetes 120:340–350Google Scholar
  13. 13.
    Shimokawa I, Trindade LS (2010) Dietary restriction and aging in rodents: a current view on its molecular mechanisms. Aging Dis 1:89–107Google Scholar
  14. 14.
    Xu BL, Wang R, Ma LN, Dong W, Zhao ZW, Zhang JS, Wang YL, Zhang X (2015) Effects of caloric intake on learning and memory function in juvenile C57BL/6J mice. Biomed Res Int 2015:759803Google Scholar
  15. 15.
    Smialowska M, Domin H, Zieba B, Kozniewska E, Michalik R, Piotrowski P, Kajta M (2009) Neuroprotective effects of neuropeptide Y-Y2 and Y5 receptor agonists in vitro and in vivo. Neuropeptides 43:235–249Google Scholar
  16. 16.
    Decressac M, Pain S, Chabeauti PY, Frangeul L, Thiriet N, Herzog H, Vergote J, Chalon S, Jaber M, Gaillard A (2012) Neuroprotection by neuropeptide Y in cell and animal models of Parkinson’s disease. Neurobiol Aging 33:2125–2137Google Scholar
  17. 17.
    Minor RK, Chang JW, de Cabo R (2009) Hungry for life: how the arcuate nucleus and neuropeptide Y may play a critical role in mediating the benefits of calorie restriction. Mol Cell Endocrinol 299:79–88Google Scholar
  18. 18.
    Karasek M (2004) Melatonin, human aging, and age-related diseases. Exp Gerontol 39:1723–1729Google Scholar
  19. 19.
    Karasek M (2007) Does melatonin play a role in aging processes? J Physiol Pharmacol 58:105–113Google Scholar
  20. 20.
    Gutierrez-Cuesta J, Tajes M, Jimenez A, Coto-Montes A, Camins A, Pallas M (2008) Evaluation of potential pro-survival pathways regulated by melatonin in a murine senescence model. J Pineal Res 45:497–505Google Scholar
  21. 21.
    Tajes M, Gutierrez-Cuesta J, Ortuno-Sahagun D, Camins A, Pallas M (2009) Anti-aging properties of melatonin in an in vitro murine senescence model: involvement of the sirtuin 1 pathway. J Pineal Res 47:228–237Google Scholar
  22. 22.
    Cristofol R, Porquet D, Corpas R, Coto-Montes A, Serret J, Camins A, Pallas M, Sanfeliu C (2012) Neurons from senescence-accelerated SAMP8 mice are protected against frailty by the sirtuin 1 promoting agents melatonin and resveratrol. J Pineal Res 52:271–281Google Scholar
  23. 23.
    Chang HM, Wu UI, Lan CT (2009) Melatonin preserves longevity protein (sirtuin 1) expression in the hippocampus of total sleep-deprived rats. J Pineal Res 47:211–220Google Scholar
  24. 24.
    Kong PJ, Byun JS, Lim SY, Lee JJ, Hong SJ, Kwon KJ, Kim SS (2008) Melatonin induces Akt phosphorylation through melatonin receptor- and PI3K-dependent pathways in primary astrocytes. Korean J Physiol Pharmacol 12:37–41Google Scholar
  25. 25.
    Koh PO (2008) Melatonin prevents the injury-induced decline of Akt/forkhead transcription factors phosphorylation. J Pineal Res 45:199–203Google Scholar
  26. 26.
    Park S, Fujishita C, Komatsu T, Kim SE, Chiba T, Mori R, Shimokawa I (2014) NPY antagonism reduces adiposity and attenuates age-related imbalance of adipose tissue metabolism. FASEB J 28:5337–5348Google Scholar
  27. 27.
    Hayashi H, Yamaza H, Komatsu T, Park S, Chiba T, Higami Y, Nagayasu T (2008) Calorie restriction minimizes activation of insulin signaling in response to glucose: potential involvement of the growth hormone-insulin-like growth factor 1 axis. Exp Gerontol 43:827–832Google Scholar
  28. 28.
    Bannon AW, Seda J, Carmouche M, Francis JM, Norman MH, Karbon B, McCaleb ML (2000) Behavioral characterization of neuropeptide Y knockout mice. Brain Res 868:79–87Google Scholar
  29. 29.
    Chiba T, Tamashiro Y, Park D, Kusudo T, Fujie R, Komatsu T, Kim SE, Park S, Hayashi H, Mori R, Yamashita H, Chung HY, Shimokawa I (2014) A key role for neuropeptide Y in lifespan extension and cancer suppression via dietary restriction. Sci Rep 4:4517Google Scholar
  30. 30.
    Park SJ, Shin EJ, Min SS, An J, Li Z, Hee Chung Y, Hoon Jeong J, Bach JH, Nah SY, Kim WK, Jang CG, Kim YS, Nabeshima Y, Nabeshima T, Kim HC (2013) Inactivation of JAK2/STAT3 signaling axis and downregulation of M1 mAChR cause cognitive impairment in klotho mutant mice, a genetic model of aging. Neuropsychopharmacology 38:1426–1437Google Scholar
  31. 31.
    Shin EJ, Chung YH, Le HL, Jeong JH, Dang DK, Nam Y, Wie MB, Nah SY, Nabeshima Y, Nabeshima T, Kim HC (2014) Melatonin attenuates memory impairment induced by Klotho gene deficiency via interactive signaling between MT2 receptor, ERK, and Nrf2-related antioxidant potential. Int J Neuropsychopharmacol. doi:
  32. 32.
    Tan DX, Manchester LC, Sainz RM, Mayo JC, Leon J, Hardeland R, Poeggeler B, Reiter RJ (2005) Interactions between melatonin and nicotinamide nucleotide: NADH preservation in cells and in cell-free systems by melatonin. J Pineal Res 39:185–194Google Scholar
  33. 33.
    Graff J, Kahn M, Samiei A, Gao J, Ota KT, Rei D, Tsai LH (2013) A dietary regimen of caloric restriction or pharmacological activation of SIRT1 to delay the onset of neurodegeneration. J Neurosci 33:8951–8960Google Scholar
  34. 34.
    Turan B, Tuncay E, Vassort G (2012) Resveratrol and diabetic cardiac function: focus on recent in vitro and in vivo studies. J Bioenerg Biomembr 44:281–296Google Scholar
  35. 35.
    Liu D, Pitta M, Mattson MP (2008) Preventing NAD(+) depletion protects neurons against excitotoxicity: bioenergetic effects of mild mitochondrial uncoupling and caloric restriction. Ann NY Acad Sci 1147:275–282Google Scholar
  36. 36.
    Lu SP, Lin SJ (2010) Regulation of yeast sirtuins by NAD(+) metabolism and calorie restriction. Biochim Biophys Acta 1804:1567–1575Google Scholar
  37. 37.
    Ramis MR, Esteban S, Miralles A, Tan DX, Reiter RJ (2015) Caloric restriction, resveratrol and melatonin: role of SIRT1 and implications for aging and related-diseases. Mech Ageing Dev 146–148:28–41Google Scholar
  38. 38.
    Rahat O, Maoz N, Cohen HY (2011) Multiple pathways regulating the calorie restriction response in yeast. J Gerontol A 66:163–169Google Scholar
  39. 39.
    Anderson RM, Bitterman KJ, Wood JG, Medvedik O, Sinclair DA (2003) Nicotinamide and PNC1 govern lifespan extension by calorie restriction in Saccharomyces cerevisiae. Nature 423:181–185Google Scholar
  40. 40.
    Song J, Ke SF, Zhou CC, Zhang SL, Guan YF, Xu TY, Sheng CQ, Wang P, Miao CY (2014) Nicotinamide phosphoribosyltransferase is required for the calorie restriction-mediated improvements in oxidative stress, mitochondrial biogenesis, and metabolic adaptation. J Gerontol A Biol Sci Med Sci 69:44–57Google Scholar
  41. 41.
    Yang H, Lavu S, Sinclair DA (2006) Nampt/PBEF/Visfatin: a regulator of mammalian health and longevity? Exp Gerontol 41:718–726Google Scholar
  42. 42.
    Yamamoto T, Byun J, Zhai P, Ikeda Y, Oka S, Sadoshima J (2014) Nicotinamide mononucleotide, an intermediate of NAD+ synthesis, protects the heart from ischemia and reperfusion. PLoS ONE 9:e98972Google Scholar
  43. 43.
    Caricasole A, Copani A, Caraci F, Aronica E, Rozemuller AJ, Caruso A, Storto M, Gaviraghi G, Terstappen GC, Nicoletti F (2004) Induction of Dickkopf-1, a negative modulator of the Wnt pathway, is associated with neuronal degeneration in Alzheimer’s brain. J Neurosci 24:6021–6027Google Scholar
  44. 44.
    Chiba T, Kamei Y, Shimizu T, Shirasawa T, Katsumata A, Shiraishi L, Sugita S, Ogawa Y, Miura S, Ezaki O (2009) Overexpression of FOXO1 in skeletal muscle does not alter longevity in mice. Mech Ageing Dev 130:420–428Google Scholar
  45. 45.
    Bousiges O, Vasconcelos AP, Neidl R, Cosquer B, Herbeaux K, Panteleeva I, Loeffler JP, Cassel JC, Boutillier AL (2010) Spatial memory consolidation is associated with induction of several lysine-acetyltransferase (histone acetyltransferase) expression levels and H2B/H4 acetylation-dependent transcriptional events in the rat hippocampus. Neuropsychopharmacology 35:2521–2537Google Scholar
  46. 46.
    Wang J, Xiao X, Zhang Y, Shi D, Chen W, Fu L, Liu L, Xie F, Kang T, Huang W, Deng W (2012) Simultaneous modulation of COX-2, p300, Akt, and Apaf-1 signaling by melatonin to inhibit proliferation and induce apoptosis in breast cancer cells. J Pineal Res 53:77–90Google Scholar
  47. 47.
    Shi D, Xiao X, Wang J, Liu L, Chen W, Fu L, Xie F, Huang W, Deng W (2012) Melatonin suppresses proinflammatory mediators in lipopolysaccharide-stimulated CRL1999 cells via targeting MAPK, NF-kappaB, c/EBPbeta, and p300 signaling. J Pineal Res 53:154–165Google Scholar
  48. 48.
    Anacker C, Zunszain PA, Cattaneo A, Carvalho LA, Garabedian MJ, Thuret S, Price J, Pariante CM (2011) Antidepressants increase human hippocampal neurogenesis by activating the glucocorticoid receptor. Mol Psychiatry 16:738–750Google Scholar
  49. 49.
    Andreu Z, Khan MA, Gonzalez-Gomez P, Negueruela S, Hortiguela R, San Emeterio J, Ferron SR, Martinez G, Vidal A, Farinas I, Lie DC, Mira H (2015) The cyclin-dependent kinase inhibitor p27 kip1 regulates radial stem cell quiescence and neurogenesis in the adult hippocampus. Stem Cells 33:219–229Google Scholar
  50. 50.
    Heine VM, Maslam S, Joels M, Lucassen PJ (2004) Increased P27KIP1 protein expression in the dentate gyrus of chronically stressed rats indicates G1 arrest involvement. Neuroscience 129:593–601Google Scholar
  51. 51.
    Berschneider B, Konigshoff M (2011) WNT1 inducible signaling pathway protein 1 (WISP1): a novel mediator linking development and disease. Int J Biochem Cell Biol 43:306–309Google Scholar
  52. 52.
    Wang S, Zhong Chong Z, Chen Shang Y, Maiese K (2013) WISP1 neuroprotection requires FoxO3a post-translational modulation with autoregulatory control of SIRT1. Curr Neurovasc Res 10:54–69Google Scholar
  53. 53.
    Su F, Overholtzer M, Besser D, Levine AJ (2002) WISP-1 attenuates p53-mediated apoptosis in response to DNA damage through activation of the Akt kinase. Genes Dev 16:46–57Google Scholar
  54. 54.
    Sequea DA, Sharma N, Arias EB, Cartee GD (2013) Greater filamin C, GSK3alpha, and GSK3beta serine phosphorylation in insulin-stimulated isolated skeletal muscles of calorie restricted 24 month-old rats. Mech Ageing Dev 134:60–63Google Scholar
  55. 55.
    Tajes Orduna M, Pelegri Gabalda C, Vilaplana Hortensi J, Pallas Lliberia M, Camins Espuny A (2009) An evaluation of the neuroprotective effects of melatonin in an in vitro experimental model of age-induced neuronal apoptosis. J Pineal Res 46:262–267Google Scholar
  56. 56.
    Gutierrez-Cuesta J, Sureda FX, Romeu M, Canudas AM, Caballero B, Coto-Montes A, Camins A, Pallas M (2007) Chronic administration of melatonin reduces cerebral injury biomarkers in SAMP8. J Pineal Res 42:394–402Google Scholar
  57. 57.
    Peng CX, Hu J, Liu D, Hong XP, Wu YY, Zhu LQ, Wang JZ (2013) Disease-modified glycogen synthase kinase-3beta intervention by melatonin arrests the pathology and memory deficits in an Alzheimer’s animal model. Neurobiol Aging 34:1555–1563Google Scholar
  58. 58.
    Kwon KJ, Kim HJ, Shin CY, Han SH (2010) Melatonin potentiates the neuroprotective properties of resveratrol against beta-amyloid-induced neurodegeneration by modulating AMP-activated protein kinase pathways. J Clin Neurol 6:127–137Google Scholar
  59. 59.
    Dietrich MO, Antunes C, Geliang G, Liu ZW, Borok E, Nie Y, Xu AW, Souza DO, Gao Q, Diano S, Gao XB, Horvath TL (2010) Agrp neurons mediate Sirt1’s action on the melanocortin system and energy balance: roles for Sirt1 in neuronal firing and synaptic plasticity. J Neurosci 30:11815–11825Google Scholar
  60. 60.
    Simonneaux V, Ribelayga C (2003) Generation of the melatonin endocrine message in mammals: a review of the complex regulation of melatonin synthesis by norepinephrine, peptides, and other pineal transmitters. Pharmacol Rev 55:325–395Google Scholar
  61. 61.
    Díaz E, Debeljuk L, Arce A, Esquifino A, Díaz B (2000) Prenatal melatonin exposure affects luteinizing hormone and hypothalamic and striatal neuropeptide Y in the male rat offspring. Neurosci Lett 292:143–146Google Scholar
  62. 62.
    Aydin M, Canpolat S, Kuloglu T, Yasar A, Colakoglu N, Kelestimur H (2008) Effects of pinealectomy and exogenous melatonin on ghrelin and peptide YY in gastrointestinal system and neuropeptide Y in hypothalamic arcuate nucleus: immunohistochemical studies in male rats. Regul Pept 146:197–203Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2017

Authors and Affiliations

  • Anorut Jenwitheesuk
    • 1
  • Seongjoon Park
    • 2
  • Prapimpun Wongchitrat
    • 3
  • Jiraporn Tocharus
    • 4
  • Sujira Mukda
    • 1
  • Isao Shimokawa
    • 2
    Email author
  • Piyarat Govitrapong
    • 1
    • 5
    • 6
    Email author
  1. 1.Research Center for Neuroscience, Institute of Molecular BiosciencesMahidol UniversitySalayaThailand
  2. 2.Department of PathologyNagasaki University School of Medicine and Graduate School of Biomedical SciencesNagasakiJapan
  3. 3.Center for Research and Innovation, Faculty of Medical TechnologyMahidol UniversitySalayaThailand
  4. 4.Department of Physiology, Faculty of MedicineChiang Mai UniversityChiang MaiThailand
  5. 5.Center for Neuroscience and Department of Pharmacology, Faculty of ScienceMahidol UniversitySalayaThailand
  6. 6.Chulabhorn Graduate InstituteChulabhorn Royal AcademyBangkokThailand

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