Skip to main content
Top
Published in: BMC Complementary Medicine and Therapies 1/2013

Open Access 01-12-2013 | Research article

Neuritogenic effect of standardized extract of Centella asiatica ECa233 on human neuroblastoma cells

Authors: Oraphan Wanakhachornkrai, Varisa Pongrakhananon, Preedakorn Chunhacha, Aree Wanasuntronwong, Anusara Vattanajun, Boonyong Tantisira, Pithi Chanvorachote, Mayuree H Tantisira

Published in: BMC Complementary Medicine and Therapies | Issue 1/2013

Login to get access

Abstract

Background

In order to gain insight into neuroprotective effects of ECa 233, a standardized extract of Centella asiatica, previously demonstrated in animal models of memory impairment induced by transient global ischemia or intracerebroventricular injection of β-amyloid, the effect of ECa 233 on neurite outgrowth of human IMR-32 neuroblastoma cell line was investigated.

Methods

Cells were seeded and incubated with various concentrations of ECa 233. Morphometric analysis was carried out by a measurement of the longest neurite growth of cells at 24 and 48 h. Contributing signaling pathways possibly involved were subsequently elucidated by western blot analysis.

Results

While ECa 233 had only limited effects on cell viability, it significantly enhanced neurite outgrowth of IMR-32 cells at the concentrations of 1–100 μg/ml. Western blot analysis revealed that ECa 233 significantly upregulated the level of activated ERK1/2 and Akt of the treated cells suggesting their involvement in the neuritogenic effect observed, which was subsequently verified by the finding that an addition of their respective inhibitors could reverse the effect of ECa 233 on these cells.

Conclusions

The present study clearly demonstrated neurite outgrowth promoting activity of ECa 233. ERK1/2 and Akt signaling pathways seemed to account for the neurotrophic effect observed. In conjunction with in vivo neuroprotective effect of ECa 233 previously reported, the results obtained support further development of ECa 233 for clinical use in neuronal injury or neurodegenerative diseases.
Appendix
Available only for authorised users
Literature
1.
go back to reference Plachez C, Richards LJ: Mechanisms of axon guidance in the developing nervous system. Curr Top Dev Biol. 2005, 69: 267-346.CrossRefPubMed Plachez C, Richards LJ: Mechanisms of axon guidance in the developing nervous system. Curr Top Dev Biol. 2005, 69: 267-346.CrossRefPubMed
2.
go back to reference Kaplan D, Miller F: Neurotrophin signaling transduction in the nervous system. Curr Opin Neurobiol. 2000, 10: 381-391. 10.1016/S0959-4388(00)00092-1.CrossRefPubMed Kaplan D, Miller F: Neurotrophin signaling transduction in the nervous system. Curr Opin Neurobiol. 2000, 10: 381-391. 10.1016/S0959-4388(00)00092-1.CrossRefPubMed
3.
go back to reference Hur J, Lee P, Kim H, Kang I, Lee KR, Kim SY: (−)-3,5-Dicaffeoyl-muco-quinic acid isolated from Aster scaber contributes to the differentiation of PC12 cells: through tyrosine kinase cascade signaling. Biochem and Biophys Res Commun. 2004, 313: 948-953. 10.1016/j.bbrc.2003.11.178.CrossRef Hur J, Lee P, Kim H, Kang I, Lee KR, Kim SY: (−)-3,5-Dicaffeoyl-muco-quinic acid isolated from Aster scaber contributes to the differentiation of PC12 cells: through tyrosine kinase cascade signaling. Biochem and Biophys Res Commun. 2004, 313: 948-953. 10.1016/j.bbrc.2003.11.178.CrossRef
4.
go back to reference Zhai H, Nakade K, Oda M, Mitsumoto Y, Akagi M, Sakurai J, Fukuyama Y: Honokiol-induced neurite outgrowth promotion depends on activation of extracellular signal-regulated kinases (ERK1/2). Eur J Pharmacol. 2005, 516: 112-117. 10.1016/j.ejphar.2005.04.035.CrossRefPubMed Zhai H, Nakade K, Oda M, Mitsumoto Y, Akagi M, Sakurai J, Fukuyama Y: Honokiol-induced neurite outgrowth promotion depends on activation of extracellular signal-regulated kinases (ERK1/2). Eur J Pharmacol. 2005, 516: 112-117. 10.1016/j.ejphar.2005.04.035.CrossRefPubMed
5.
go back to reference Read DE, Gorman AM: Involvement of Akt in neurite outgrowth. Cell Mol Life Sci. 2009, 66: 2975-2984. 10.1007/s00018-009-0057-8.CrossRefPubMed Read DE, Gorman AM: Involvement of Akt in neurite outgrowth. Cell Mol Life Sci. 2009, 66: 2975-2984. 10.1007/s00018-009-0057-8.CrossRefPubMed
6.
go back to reference Takahashi K, Piao S, Yamatani H, Du B, Yin L, Ohta T, Kawagoe J, Takata K, Tsutsumi S, Kurachi H: Estrogen induces neurite outgrowth via Rho family GTPases in neuroblastoma cells. Mol Cell Neurosci. 2011, 48: 217-224. 10.1016/j.mcn.2011.08.002.CrossRefPubMed Takahashi K, Piao S, Yamatani H, Du B, Yin L, Ohta T, Kawagoe J, Takata K, Tsutsumi S, Kurachi H: Estrogen induces neurite outgrowth via Rho family GTPases in neuroblastoma cells. Mol Cell Neurosci. 2011, 48: 217-224. 10.1016/j.mcn.2011.08.002.CrossRefPubMed
7.
go back to reference Labelle C, Leclerc N: Exogenouse BDNF, NT-3 and NT-4 differentially regulate neurite outgrowth in cultured hippocampal neurons. Brain Res Dev Brain Res. 2000, 123: 1-11. 10.1016/S0165-3806(00)00069-9.CrossRefPubMed Labelle C, Leclerc N: Exogenouse BDNF, NT-3 and NT-4 differentially regulate neurite outgrowth in cultured hippocampal neurons. Brain Res Dev Brain Res. 2000, 123: 1-11. 10.1016/S0165-3806(00)00069-9.CrossRefPubMed
8.
go back to reference Chao MV, Rajagopal R, Lee FS: Neurotrophin signaling in health and disease. Clin Sci (Lond). 2006, 110: 167-173. 10.1042/CS20050163.CrossRef Chao MV, Rajagopal R, Lee FS: Neurotrophin signaling in health and disease. Clin Sci (Lond). 2006, 110: 167-173. 10.1042/CS20050163.CrossRef
9.
go back to reference Küenzi P, Kiefer S, Koryalina A, Hamburger M: Promotion of cell death or neurite outgrowth in PC-12 and N2a cells by the fungal alkaloid militarinone A depends on basal expression of p53. Apoptosis. 2008, 13: 364-376. 10.1007/s10495-008-0185-x.CrossRefPubMed Küenzi P, Kiefer S, Koryalina A, Hamburger M: Promotion of cell death or neurite outgrowth in PC-12 and N2a cells by the fungal alkaloid militarinone A depends on basal expression of p53. Apoptosis. 2008, 13: 364-376. 10.1007/s10495-008-0185-x.CrossRefPubMed
10.
go back to reference Wang X, Wang Z, Yao Y, Li J, Zhang X, Li C, Cheng Y, Ding G, Liu L, Ding Z: Essential role of ERK activation in neurite outgrowth induced by α-lipoic acid. Biochim Biophys Acta. 1813, 2011: 827-838. Wang X, Wang Z, Yao Y, Li J, Zhang X, Li C, Cheng Y, Ding G, Liu L, Ding Z: Essential role of ERK activation in neurite outgrowth induced by α-lipoic acid. Biochim Biophys Acta. 1813, 2011: 827-838.
11.
go back to reference Horner PJ, Gage FH: Regenerating the central nervous system. Nat. 2000, 407: 963-970. 10.1038/35039559.CrossRef Horner PJ, Gage FH: Regenerating the central nervous system. Nat. 2000, 407: 963-970. 10.1038/35039559.CrossRef
12.
go back to reference Soumyanath A, Zhong YP, Gold SA, Yu X, Koop DR, Bourdette D, Gold BG: Centella asiatica accelerates nerve regeneration upon oral administration and contains multiple active fractions increasing neurite elongation in-vitro. J Pharm Pharmacol. 2005, 57: 1221-1229. 10.1211/jpp.57.9.0018.CrossRefPubMed Soumyanath A, Zhong YP, Gold SA, Yu X, Koop DR, Bourdette D, Gold BG: Centella asiatica accelerates nerve regeneration upon oral administration and contains multiple active fractions increasing neurite elongation in-vitro. J Pharm Pharmacol. 2005, 57: 1221-1229. 10.1211/jpp.57.9.0018.CrossRefPubMed
13.
go back to reference Maier IC, Schweb ME: Sprouting, regeneration and circuit formation in the injured spinal cord: factors and activity. Philos Trans R Soc Lond B Biol Sci. 2006, 61: 1611-1634.CrossRef Maier IC, Schweb ME: Sprouting, regeneration and circuit formation in the injured spinal cord: factors and activity. Philos Trans R Soc Lond B Biol Sci. 2006, 61: 1611-1634.CrossRef
14.
go back to reference Brinkhaus B, Lindner M, Schuppan D, Hahn EG: Chemical, pharmacological and clinical profile of the East Asian medical plant Centella asiatica. Phytomedicine. 2000, 7: 427-448. 10.1016/S0944-7113(00)80065-3.CrossRefPubMed Brinkhaus B, Lindner M, Schuppan D, Hahn EG: Chemical, pharmacological and clinical profile of the East Asian medical plant Centella asiatica. Phytomedicine. 2000, 7: 427-448. 10.1016/S0944-7113(00)80065-3.CrossRefPubMed
15.
go back to reference Inamdar PK, Yeole RD, Ghogare AB, de Souza NJ: Determination of biologically active constituents in Centella asiatica. J Chromatography A. 1996, 742: 127-130. 10.1016/0021-9673(96)00237-3.CrossRef Inamdar PK, Yeole RD, Ghogare AB, de Souza NJ: Determination of biologically active constituents in Centella asiatica. J Chromatography A. 1996, 742: 127-130. 10.1016/0021-9673(96)00237-3.CrossRef
16.
go back to reference Wanasuntronwong A, Tantisira MH, Tantisira B, Watanabe H: Anxiolytic effects of standardized extract of Centella asiatica (ECa 233) after chronic immobilization stress in mice. J Ethnopharmacol. 2012, 143: 579-585. 10.1016/j.jep.2012.07.010.CrossRefPubMed Wanasuntronwong A, Tantisira MH, Tantisira B, Watanabe H: Anxiolytic effects of standardized extract of Centella asiatica (ECa 233) after chronic immobilization stress in mice. J Ethnopharmacol. 2012, 143: 579-585. 10.1016/j.jep.2012.07.010.CrossRefPubMed
17.
go back to reference Tantisira MH, Tantisira B, Patarapanich C, Suttisri R, Luangcholatan S, Mingmalailak S, Wanasuntronwong A, Saifah E: Effects of standardized extract of Centella asiatica ECa 233 on learning and memory impairment induced by transient bilateral common carotid artery occlusion in mice. Thai J Pharmacol. 2010, 32: 22-33. Tantisira MH, Tantisira B, Patarapanich C, Suttisri R, Luangcholatan S, Mingmalailak S, Wanasuntronwong A, Saifah E: Effects of standardized extract of Centella asiatica ECa 233 on learning and memory impairment induced by transient bilateral common carotid artery occlusion in mice. Thai J Pharmacol. 2010, 32: 22-33.
18.
go back to reference Kam-eg A, Tantisira B, Tantisira MH: Preliminary study on effects of standardized extract of Centella asiatica, Eca 233, on deficit of learning and memory induced by an intracerebroventricular injection of β-amyloid peptide in mice. Thai J Pharmacol. 2009, 31: 79-82. Kam-eg A, Tantisira B, Tantisira MH: Preliminary study on effects of standardized extract of Centella asiatica, Eca 233, on deficit of learning and memory induced by an intracerebroventricular injection of β-amyloid peptide in mice. Thai J Pharmacol. 2009, 31: 79-82.
19.
go back to reference Fukuyama Y, Nakade K, Minoshima Y, Yokoyama R, Zhai H, Mitsumoto Y: Neurotrophic activity of honokiol on the cultures of fetal rat cortical neurons. Bioorg Med Chem Lett. 2002, 12: 1163-1166. 10.1016/S0960-894X(02)00112-9.CrossRefPubMed Fukuyama Y, Nakade K, Minoshima Y, Yokoyama R, Zhai H, Mitsumoto Y: Neurotrophic activity of honokiol on the cultures of fetal rat cortical neurons. Bioorg Med Chem Lett. 2002, 12: 1163-1166. 10.1016/S0960-894X(02)00112-9.CrossRefPubMed
20.
go back to reference Hur J, Lee P, Moon E, Kang I, Kim SH, Oh MH, Kim Y: Neurite outgrowth induced by spicatoside A, a steroidal saponin, via the tyrosine kinase A receptor pathway. Eur J Pharmacol. 2009, 620: 9-15. 10.1016/j.ejphar.2009.08.016.CrossRefPubMed Hur J, Lee P, Moon E, Kang I, Kim SH, Oh MH, Kim Y: Neurite outgrowth induced by spicatoside A, a steroidal saponin, via the tyrosine kinase A receptor pathway. Eur J Pharmacol. 2009, 620: 9-15. 10.1016/j.ejphar.2009.08.016.CrossRefPubMed
21.
go back to reference Taylor JLS, Rabe T, McGaw LJ, Jäger AK, Staden Van J: Toward the scientific validation of traditional medicinal plants. Plant Growth Regul. 2001, 34: 23-37. 10.1023/A:1013310809275.CrossRef Taylor JLS, Rabe T, McGaw LJ, Jäger AK, Staden Van J: Toward the scientific validation of traditional medicinal plants. Plant Growth Regul. 2001, 34: 23-37. 10.1023/A:1013310809275.CrossRef
22.
go back to reference Eckert GP: Traditional used plants against cognitive decline and alzheimer disease. Front Pharmcol. 2010, 1: 1-10. Eckert GP: Traditional used plants against cognitive decline and alzheimer disease. Front Pharmcol. 2010, 1: 1-10.
23.
go back to reference Wattanatorn J, Mator L, Muchimapura S, Tongun T, Pasuriwong O, Piyawatkul N, Yimtae K, Sripanidkulchai B, Singkhoraard J: Positive modulation of cognition and mood in the healthy elderly volunteer following the administration of Centella asiatica. J Ethnopharmacol. 2008, 116: 325-332. 10.1016/j.jep.2007.11.038.CrossRef Wattanatorn J, Mator L, Muchimapura S, Tongun T, Pasuriwong O, Piyawatkul N, Yimtae K, Sripanidkulchai B, Singkhoraard J: Positive modulation of cognition and mood in the healthy elderly volunteer following the administration of Centella asiatica. J Ethnopharmacol. 2008, 116: 325-332. 10.1016/j.jep.2007.11.038.CrossRef
24.
go back to reference Rao SB, Chetana M, Uma Devi P: Centella asiatica treatment during postnatal period enhances learning and memory in mice. Physiol Behav. 2005, 86: 449-457. 10.1016/j.physbeh.2005.07.019.CrossRefPubMed Rao SB, Chetana M, Uma Devi P: Centella asiatica treatment during postnatal period enhances learning and memory in mice. Physiol Behav. 2005, 86: 449-457. 10.1016/j.physbeh.2005.07.019.CrossRefPubMed
25.
go back to reference Mohandas Rao KG, Muddanna Rao S, Gurumadhva Rao S: Centella asiatica (L.) Leaf extract treatment during the growth spurt period enhances hippocampal CA3 neuronal dendritic arborization in rats. Evid Based Complement Alternat Med. 2006, 3: 349-357. 10.1093/ecam/nel024.CrossRefPubMedPubMedCentral Mohandas Rao KG, Muddanna Rao S, Gurumadhva Rao S: Centella asiatica (L.) Leaf extract treatment during the growth spurt period enhances hippocampal CA3 neuronal dendritic arborization in rats. Evid Based Complement Alternat Med. 2006, 3: 349-357. 10.1093/ecam/nel024.CrossRefPubMedPubMedCentral
26.
go back to reference Mohandas Rao KG, Muddanna Rao S, Gurumadhva Rao S: Enhancement of amygdaloid neuronal dendritic arboritezation by fresh leaf juice of Centella asiatica (Linn) during growth spurt period in rats. Evid Based Complement Alternat Med. 2009, 6: 575-611.CrossRef Mohandas Rao KG, Muddanna Rao S, Gurumadhva Rao S: Enhancement of amygdaloid neuronal dendritic arboritezation by fresh leaf juice of Centella asiatica (Linn) during growth spurt period in rats. Evid Based Complement Alternat Med. 2009, 6: 575-611.CrossRef
27.
go back to reference Gadahad MR, Rao M, Rao G: Enhancement of hippocampal CA3 neuronal dendritic arboritezation by Centella asiatica (Linn) fresh leaf extract in adult rats. J Chin Med Assoc. 2008, 71: 6-13. 10.1016/S1726-4901(08)70066-2.CrossRefPubMed Gadahad MR, Rao M, Rao G: Enhancement of hippocampal CA3 neuronal dendritic arboritezation by Centella asiatica (Linn) fresh leaf extract in adult rats. J Chin Med Assoc. 2008, 71: 6-13. 10.1016/S1726-4901(08)70066-2.CrossRefPubMed
28.
go back to reference Boglari G, Szeberenyi J: Nerve growth factor in combination with second messenger analogues causes neuronal differentiation of PC12 cells expressing a dominant inhibitory Ras protein without inducing activation of extracellular signal- regulated kinases. Eur J Neurosci. 2001, 14: 1445-1454. 10.1046/j.0953-816x.2001.01787.x.CrossRefPubMed Boglari G, Szeberenyi J: Nerve growth factor in combination with second messenger analogues causes neuronal differentiation of PC12 cells expressing a dominant inhibitory Ras protein without inducing activation of extracellular signal- regulated kinases. Eur J Neurosci. 2001, 14: 1445-1454. 10.1046/j.0953-816x.2001.01787.x.CrossRefPubMed
29.
go back to reference Tsuji M, Inanami O, Kuwabara M: Induction of neurite outgrowth in PC12 cells by alpha-phenyl-N-tert-butylnitron through activation of protein kinase C and the ras-extracellular signal-regulated kinase pathway. J Biol Chem. 2001, 276: 32779-32785. 10.1074/jbc.M101403200.CrossRefPubMed Tsuji M, Inanami O, Kuwabara M: Induction of neurite outgrowth in PC12 cells by alpha-phenyl-N-tert-butylnitron through activation of protein kinase C and the ras-extracellular signal-regulated kinase pathway. J Biol Chem. 2001, 276: 32779-32785. 10.1074/jbc.M101403200.CrossRefPubMed
30.
go back to reference Aletsee C, Beros A, Mullen L, Palacios S, Pak K, Dazert S, Ryan AF: Ras/MEK but not p38 signaling mediates NT-3-induced neurite extension from spiral ganglion neurons. J Assoc Res Otolaryngol. 2001, 2: 377-387. 10.1007/s10162001000086.CrossRefPubMedPubMedCentral Aletsee C, Beros A, Mullen L, Palacios S, Pak K, Dazert S, Ryan AF: Ras/MEK but not p38 signaling mediates NT-3-induced neurite extension from spiral ganglion neurons. J Assoc Res Otolaryngol. 2001, 2: 377-387. 10.1007/s10162001000086.CrossRefPubMedPubMedCentral
31.
go back to reference Bosco A, Linden R: BDNF and NT-4 differentially modulate neurite outgrowth in developing retinal ganglion cells. J Neurosci Res. 1999, 15: 759-769.CrossRef Bosco A, Linden R: BDNF and NT-4 differentially modulate neurite outgrowth in developing retinal ganglion cells. J Neurosci Res. 1999, 15: 759-769.CrossRef
32.
go back to reference Price RD, Yamaji T, Matsuoka N: FK506 potentiates NGF-induced neurite outgrowth via the Ras/Raf/MAP kinase pathway. J Pharmacol. 2003, 140: 825-829. Price RD, Yamaji T, Matsuoka N: FK506 potentiates NGF-induced neurite outgrowth via the Ras/Raf/MAP kinase pathway. J Pharmacol. 2003, 140: 825-829.
33.
go back to reference Gold BG, Zhong YP: FK506 requires stimulation of the extracellular signal regulated kinase 1/2 and the steroid receptor chaperone protein p23 for neurite elongation. Neurosignals. 2004, 13: 122-129. 10.1159/000076565.CrossRefPubMed Gold BG, Zhong YP: FK506 requires stimulation of the extracellular signal regulated kinase 1/2 and the steroid receptor chaperone protein p23 for neurite elongation. Neurosignals. 2004, 13: 122-129. 10.1159/000076565.CrossRefPubMed
34.
go back to reference Yamazaki M, Chiba K, Mohri T, Hatanaka H: Cyclic GMP-dependent neurite outgrowth by genipin and nerve growth factor in PC12h cells. Eur J Pharmacol. 2004, 488: 35-43. 10.1016/j.ejphar.2004.02.009.CrossRefPubMed Yamazaki M, Chiba K, Mohri T, Hatanaka H: Cyclic GMP-dependent neurite outgrowth by genipin and nerve growth factor in PC12h cells. Eur J Pharmacol. 2004, 488: 35-43. 10.1016/j.ejphar.2004.02.009.CrossRefPubMed
35.
go back to reference Rudakewich M, Ba F, Benishin CG: Neurotrophic and neuroprotective action of Ginsenosides Rb1 and Rg1. Planta Med. 2000, 67: 533-537.CrossRef Rudakewich M, Ba F, Benishin CG: Neurotrophic and neuroprotective action of Ginsenosides Rb1 and Rg1. Planta Med. 2000, 67: 533-537.CrossRef
36.
go back to reference Chivapat S, Chavalittumrong P, Tantisira MH: Acute and sub chronic toxicity studies of a standardized extract of Centella asiatica ECa 233. Thai J Pharm Sci. 2011, 35: 55-64. Chivapat S, Chavalittumrong P, Tantisira MH: Acute and sub chronic toxicity studies of a standardized extract of Centella asiatica ECa 233. Thai J Pharm Sci. 2011, 35: 55-64.
Metadata
Title
Neuritogenic effect of standardized extract of Centella asiatica ECa233 on human neuroblastoma cells
Authors
Oraphan Wanakhachornkrai
Varisa Pongrakhananon
Preedakorn Chunhacha
Aree Wanasuntronwong
Anusara Vattanajun
Boonyong Tantisira
Pithi Chanvorachote
Mayuree H Tantisira
Publication date
01-12-2013
Publisher
BioMed Central
Published in
BMC Complementary Medicine and Therapies / Issue 1/2013
Electronic ISSN: 2662-7671
DOI
https://doi.org/10.1186/1472-6882-13-204

Other articles of this Issue 1/2013

BMC Complementary Medicine and Therapies 1/2013 Go to the issue