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Cellular and Molecular Neurobiology

, Volume 34, Issue 6, pp 913–923 | Cite as

Bilobalide Induces Neuronal Differentiation of P19 Embryonic Carcinoma Cells via Activating Wnt/β-Catenin Pathway

  • Mei Liu
  • Jingjing Guo
  • Juan Wang
  • Luyong Zhang
  • Tao PangEmail author
  • Hong LiaoEmail author
Original Research

Abstract

Bilobalide, a natural product extracted from Ginkgo biloba leaf, is known to exhibit a number of pharmacological activities. So far, whether it could affect embryonic stem cell differentiation is still unknown. The main aim of this study was to investigate the effect of bilobalide on P19 embryonic carcinoma cells differentiation and the underlying mechanisms. Our results showed that bilobalide induced P19 cells differentiation into neurons in a concentration- and time-dependent manner. We also found that bilobalide promoted neuronal differentiation through activation of Wnt/β-catenin signaling pathway. Exposure to bilobalide increased inactive GSK-3β phosphorylation, further induced the nuclear accumulation of β-catenin, and also up-regulated the expression of Wnt ligands Wnt1 and Wnt7a. Neuronal differentiation induced by bilobalide was totally abolished by XAV939, an inhibitor of Wnt/β-catenin pathway. These results revealed a novel role of bilobalide in neuronal differentiation from P19 embryonic cells acting through Wnt/β-catenin signaling pathway, which would provide a better insight into the beneficial effects of bilobalide in brain diseases.

Keywords

Bilobalide Neuronal differentiation P19 embryonic carcinoma cells Wnt/β-catenin signaling 

Abbreviations

Amyloid β-peptide

ADDL

Amyloid-β derived diffusible ligand

APC

Adenomatous polyposis coli

bHLH protein

Basic helix-loop-helix protein

β-NGF

β-Nerve growth factor

BSA

Bovine serum albumin

CNPase

2′,3′-Cyclic-nucleotide 3′-phosphodiesterase

CREB

Cyclic-AMP response element binding protein

DMSO

Dimethyl sulfoxide

ESCs

Embryonic stem cells

GFAP

Glial fibrillary acidic protein

Gli

Glioma-associated oncogene family zinc finger

GSK-3β

Glycogen synthase kinase-3 beta

Ngn1

Neurogenin1

P19 EC cells

P19 embryonic carcinoma cells

RA

Retinoid acid

TCF/LEF

T-cell factor/lymphoid enhancer binding factor

Notes

Acknowledgments

This work was supported by National Natural Science Foundation of China (81271338, 81070967), the Specialized Research Fund for the Doctoral Program of Higher Education of China (20130096110011), Natural Science Foundation of Jiangsu Province (BK20130653), the Fundamental Research Funds for the Central Universities (JKZD2013006), and 2011’ Program for Excellent Scientific and Technological Innovation Team of Jiangsu Higher Education.

Conflict of interest

The authors declare no conflicts of interest.

References

  1. Ahlemeyer B, Krieglstein J (2003) Pharmacological studies supporting the therapeutic use of Ginkgo biloba extract for Alzheimer’s disease. Pharmacopsychiatry 36(Suppl 1):S8–S14PubMedGoogle Scholar
  2. Bain G, Ray WJ, Yao M, Gottlieb DI (1996) Retinoic acid promotes neural and represses mesodermal gene expression in mouse embryonic stem cells in culture. Biochem Biophys Res Commun 223:691–694PubMedCrossRefGoogle Scholar
  3. Budnik V, Salinas PC (2011) Wnt signaling during synaptic development and plasticity. Curr Opin Neurobiol 21:151–159PubMedCentralPubMedCrossRefGoogle Scholar
  4. Chatterjee SS, Kondratskaya EL, Krishtal OA (2003) Structure-activity studies with Ginkgo biloba extract constituents as receptor-gated chloride channel blockers and modulators. Pharmacopsychiatry 36(Suppl 1):S68–S77PubMedGoogle Scholar
  5. Davidson KC, Adams AM, Goodson JM, McDonald CE, Potter JC, Berndt JD, Biechele TL, Taylor RJ, Moon RT (2012) Wnt/beta-catenin signaling promotes differentiation, not self-renewal, of human embryonic stem cells and is repressed by Oct4. Proc Natl Acad SciUSA 109:4485–4490CrossRefGoogle Scholar
  6. Defeudis FV (2002) Bilobalide and neuroprotection. Pharmacol Res 46:565–568PubMedCrossRefGoogle Scholar
  7. DeFeudis FV, Drieu K (2000) Ginkgo biloba extract (EGb 761) and CNS functions: basic studies and clinical applications. Curr Drug Targets 1:25–58PubMedCrossRefGoogle Scholar
  8. DeKosky ST, Williamson JD, Fitzpatrick AL, Kronmal RA, Ives DG, Saxton JA, Lopez OL, Burke G, Carlson MC, Fried LP, Kuller LH, Robbins JA, Tracy RP, Woolard NF, Dunn L, Snitz BE, Nahin RL, Furberg CD (2008) Ginkgo biloba for prevention of dementia: a randomized controlled trial. JAMA 300:2253–2262PubMedCentralPubMedCrossRefGoogle Scholar
  9. Duchen MR (2012) Mitochondria, calcium-dependent neuronal death and neurodegenerative disease. Pflugers Arch 464:111–121PubMedCentralPubMedCrossRefGoogle Scholar
  10. Hirabayashi Y, Itoh Y, Tabata H, Nakajima K, Akiyama T, Masuyama N, Gotoh Y (2004) The Wnt/beta-catenin pathway directs neuronal differentiation of cortical neural precursor cells. Development 131:2791–2801PubMedCrossRefGoogle Scholar
  11. Huang SM, Mishina YM, Liu S, Cheung A, Stegmeier F, Michaud GA, Charlat O, Wiellette E, Zhang Y, Wiessner S, Hild M, Shi X, Wilson CJ, Mickanin C, Myer V, Fazal A, Tomlinson R, Serluca F, Shao W, Cheng H, Shultz M, Rau C, Schirle M, Schlegl J, Ghidelli S, Fawell S, Lu C, Curtis D, Kirschner MW, Lengauer C, Finan PM, Tallarico JA, Bouwmeester T, Porter JA, Bauer A, Cong F (2009) Tankyrase inhibition stabilizes axin and antagonizes Wnt signalling. Nature 461:614–620PubMedCrossRefGoogle Scholar
  12. IhI R (2012) Gingko biloba extract EGb 761: clinical data in dementia. Int Psychogeriatr 24(Suppl 1):S35–S40Google Scholar
  13. Ille F, Sommer L (2005) Wnt signaling: multiple functions in neural development. Cell Mol Life Sci 62:1100–1108PubMedCrossRefGoogle Scholar
  14. Jones-Villeneuve EM, McBurney MW, Rogers KA, Kalnins VI (1982) Retinoic acid induces embryonal carcinoma cells to differentiate into neurons and glial cells. J Cell Biol 94:253–262PubMedCrossRefGoogle Scholar
  15. Kageyama R, Ishibashi M, Takebayashi K, Tomita K (1997) bHLH transcription factors and mammalian neuronal differentiation. Int J Biochem Cell Biol 29:1389–1399PubMedCrossRefGoogle Scholar
  16. Kim S, Yoon YS, Kim JW, Jung M, Kim SU, Lee YD, Suh-Kim H (2004) Neurogenin1 is sufficient to induce neuronal differentiation of embryonal carcinoma P19 cells in the absence of retinoic acid. Cell Mol Neurobiol 24:343–356PubMedCrossRefGoogle Scholar
  17. Kim EJ, Ables JL, Dickel LK, Eisch AJ, Johnson JE (2011) Ascl1 (Mash1) defines cells with long-term neurogenic potential in subgranular and subventricular zones in adult mouse brain. PLoS ONE 6:e18472PubMedCentralPubMedCrossRefGoogle Scholar
  18. Kotakadi VS, Jin Y, Hofseth AB, Ying L, Cui X, Volate S, Chumanevich A, Wood PA, Price RL, McNeal A, Singh UP, Singh NP, Nagarkatti M, Nagarkatti PS, Matesic LE, Auclair K, Wargonich MJ, Hofseth LJ (2008) Ginkgo biloba extract EGb 761 has anti-inflammatory properties and ameliorates colitis in mice by driving effector T cell apoptosis. Carcinogenesis 29:1799–1806PubMedCentralPubMedCrossRefGoogle Scholar
  19. Kuhl SJ, Kuhl M (2013) On the role of Wnt/beta-catenin signaling in stem cells. Biochim Biophys Acta 1830:2297–2306PubMedCrossRefGoogle Scholar
  20. Lang KJ, Rathjen J, Vassilieva S, Rathjen PD (2004) Differentiation of embryonic stem cells to a neural fate: a route to re-building the nervous system? J Neurosci Res 76:184–192PubMedCrossRefGoogle Scholar
  21. Lie DC, Colamarino SA, Song HJ, Desire L, Mira H, Consiglio A, Lein ES, Jessberger S, Lansford H, Dearie AR, Gage FH (2005) Wnt signalling regulates adult hippocampal neurogenesis. Nature 437:1370–1375PubMedCrossRefGoogle Scholar
  22. Lo L, Tiveron MC, Anderson DJ (1998) MASH1 activates expression of the paired homeodomain transcription factor Phox2a, and couples pan-neuronal and subtype-specific components of autonomic neuronal identity. Development 125:609–620PubMedGoogle Scholar
  23. MacDonald BT, Tamai K, He X (2009) Wnt/beta-catenin signaling: components, mechanisms, and diseases. Dev Cell 17:9–26PubMedCentralPubMedCrossRefGoogle Scholar
  24. McBurney MW, Rogers BJ (1982) Isolation of male embryonal carcinoma cells and their chromosome replication patterns. Dev Biol 89:503–508PubMedCrossRefGoogle Scholar
  25. McBurney MW, Jones-Villeneuve EM, Edwards MK, Anderson PJ (1982) Control of muscle and neuronal differentiation in a cultured embryonal carcinoma cell line. Nature 299:165–167PubMedCrossRefGoogle Scholar
  26. Schuldiner M, Eiges R, Eden A, Yanuka O, Itskovitz-Eldor J, Goldstein RS, Benvenisty N (2001) Induced neuronal differentiation of human embryonic stem cells. Brain Res 913:201–205PubMedCrossRefGoogle Scholar
  27. Shi C, Yao Z, Xu J, Yew DT (2009) Effects of Gingko Extract (EGb761) on oxidative damage under different conditions of serum supply. J Bioenerg Biomembr 41:61–69PubMedCrossRefGoogle Scholar
  28. Shi C, Wu F, Yew DT, Xu J, Zhu Y (2010) Bilobalide prevents apoptosis through activation of the PI3K/Akt pathway in SH-SY5Y cells. Apoptosis 15:715–727PubMedCrossRefGoogle Scholar
  29. Snitz BE, O’Meara ES, Carlson MC, Arnold AM, Ives DG, Rapp SR, Saxton J, Lopez OL, Dunn LO, Sink KM, DeKosky ST (2009) Ginkgo biloba for preventing cognitive decline in older adults: a randomized trial. JAMA 23:2663–2670CrossRefGoogle Scholar
  30. Sun Y, Nadal-Vicens M, Misono S, Lin MZ, Zubiaga A, Hua X, Fan G, Greenberg ME (2001) Neurogenin promotes neurogenesis and inhibits glial differentiation by independent mechanisms. Cell 104:365–376PubMedCrossRefGoogle Scholar
  31. Tang K, Yang J, Gao X, Wang C, Liu L, Kitani H, Atsumi T, Jing N (2002) Wnt-1 promotes neuronal differentiation and inhibits gliogenesis in P19 cells. Biochem Biophys Res Commun 293:167–173PubMedCrossRefGoogle Scholar
  32. Tchantchou F, Lacor PN, Cao Z, Lao L, Hou Y, Cui C, Klein WL, Luo Y (2009) Stimulation of neurogenesis and synaptogenesis by bilobalide and quercetin via common final pathway in hippocampal neurons. J Alzheimers Dis 18:787–798PubMedGoogle Scholar
  33. Thomson JA, Itskovitz-Eldor J, Shapiro SS, Waknitz MA, Swiergiel JJ, Marshall VS, Jones JM (1998) Embryonic stem cell lines derived from human blastocysts. Science 282:1145–1147PubMedCrossRefGoogle Scholar
  34. Valvezan AJ, Klein PS (2012) GSK-3 and Wnt signaling in neurogenesis and bipolar disorder. Front Mol Neurosci 5:1PubMedCentralPubMedCrossRefGoogle Scholar
  35. Vitolo O, Gong B, Cao Z, Ishii H, Jaracz S, Nakanishi K, Arancio O, Dzyuba SV, Lefort R, Shelanski M (2009) Protection against beta-amyloid induced abnormal synaptic function and cell death by Ginkgolide. J. Neurobiol Aging 30:257–265CrossRefGoogle Scholar
  36. Wexler EM, Geschwind DH, Palmer TD (2008) Lithium regulates adult hippocampal progenitor development through canonical Wnt pathway activation. Mol Psychiatry 13:285–292PubMedCrossRefGoogle Scholar
  37. Wichterle H, Lieberam I, Porter JA, Jessell TM (2002) Directed differentiation of embryonic stem cells into motor neurons. Cell 110:385–397PubMedCrossRefGoogle Scholar
  38. Wodarz A, Nusse R (1998) Mechanisms of Wnt signaling in development. Annu Rev Cell Dev Biol 14:59–88PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Neurobiology Laboratory, National Center for Drug ScreeningChina Pharmaceutical UniversityNanjingPeople’s Republic of China
  2. 2.Key Laboratory of Drug Quality Control and Pharmacovigilance (China Pharmaceutical University), Ministry of EducationNanjingPeople’s Republic of China

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