Download PDF
Planta Med 2018; 84(16): 1165-1173
DOI: 10.1055/a-0619-5710
Biological and Pharmacological Activity
Original Papers
Georg Thieme Verlag KG Stuttgart · New York

Differences in Neuritogenic Activity and Signaling Activation of Madecassoside, Asiaticoside, and Their Aglycones in Neuro-2a cells

Nonthaneth Nalinratana
1   Department of Biochemistry and Microbiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand
,
Duangdeun Meksuriyen
2   Faculty of Pharmacy, Rangsit University, Pathum Thani, Thailand
,
Boonsri Ongpipattanakul
1   Department of Biochemistry and Microbiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand
3   Chulalongkorn University Drugs and Health Products Innovation and Promotion Center, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand
› Author Affiliations
Further Information

Publication History

received 15 January 2018
revised 06 April 2018

accepted 18 April 2018

Publication Date:
02 May 2018 (online)

Also available at
    Permissions and Reprints

Abstract

Madecassoside (MS) and asiaticoside (AS) along with their aglycones, madecassic acid (MA) and asiatic acid (AA), are considered the major neuroactive triterpenoid constituents of Centella asiatica. In this study, we aimed to compare MS, AS, MA, and AA for their neurite outgrowth activities and mechanisms in Neuro-2a cells. Immunofluorescent cell staining showed MS and AS significantly increased the percentage of neurite-bearing cells (%NBC) and the neurite length with higher potency than MA and AA. The triterpenoid glycosides induced sustained extracellular signal-regulated protein kinases 1 and 2 (ERK1/2) phosphorylation, while their aglycones activated only transient signaling of ERK1/2. Suppression of ERK1/2 activation significantly abolished not only cAMP response element-binding protein (CREB) phosphorylation but also the increment of %NBC and neurite length in MS- and AS-treated cells. Inhibition of ERK phosphorylation did not produce similar blockage of CREB activation and neurite outgrowth in MA- and AA-treated cells. On the other hand, inactivation of protein kinase B (Akt) resulted in a suppression of neurite lengthening in all studied triterpenoids. This is the first study discerning the different signaling pathways of neurite outgrowth activity induced by C. asiatica triterpenoid glycosides and aglycones. Neurite outgrowth activity of the glycosides MS and AS was found to involve the activation of sustained ERK phosphorylation leading to CREB activation, while ERK activation was not associated with MA- and AA-induced neurite outgrowth. In addition, Akt activation was evident to be more involved in neurite elongation process.

Key words

madecassoside - asiaticoside - madecassic acid - asiatic acid - neurite outgrowth - sustained ERK phosphorylation - Centella asiatica - Apiaceae
 

  • References

  • 1 Leggio MG, Mandolesi L, Federico F, Spirito F, Ricci B, Gelfo F, Petrosini L. Environmental enrichment promotes improved spatial abilities and enhanced dendritic growth in the rat. Behav Brain Res 2005; 1: 78-90
    PubMedGoogle Scholar
  • 2 Wartman BC, Holahan MR. The impact of multiple memory formation on dendritic complexity in the hippocampus and anterior cingulate cortex assessed at recent and remote time points. Front Behav Neurosci 2014; 8: 128
    PubMedGoogle Scholar
  • 3 Ramm P, Alexandrov Y, Cholewinski A, Cybuch Y, Nadon R, Soltys BJ. Automated screening of neurite outgrowth. J Biomol Screen 2003; 1: 7-18
    PubMedGoogle Scholar
  • 4 Rumalla CS, Ali Z, Weerasooriya AD, Smillie TJ, Khan IA. Two new triterpene glycosides from Centella asiatica . Planta Med 2010; 76: 1018-1021
    Article in Thieme ConnectPubMedGoogle Scholar
  • 5 Lokanathan Y, Omar N, Ahmad Puzi NN, Saim A, Hj Idrus R. Recent updates in neuroprotective and neuroregenerative potential of Centella asiatica . Malays J Med Sci 2016; 23: 4-14
    PubMedGoogle Scholar
  • 6 Tantisira M, 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
    PubMedGoogle Scholar
  • 7 Wanakhachornkrai O, Pongrakhananon V, Chunhacha P, Wanasuntronwong A, Vattanajun A, Tantisira B, Chanvorachote P, Tantisira MH. Neuritogenic effect of standardized extract of Centella asiatica ECa233 on human neuroblastoma cells. BMC Complement Altern Med 2013; 13: 204
    CrossrefPubMedGoogle Scholar
  • 8 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
    CrossrefPubMedGoogle Scholar
  • 9 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; 9: 1221-1229
    PubMedGoogle Scholar
  • 10 Jiang H, Zheng G, Lv J, Chen H, Lin J, Li Y, Fan G, Ding X. Identification of Centella asiatica s effective ingredients for inducing the neuronal differentiation. Evid Based Complement Alternat Med 2016; 2016: 9634750
    PubMedGoogle Scholar
  • 11 Gray NE, Zweig JA, Murchison C, Caruso M, Matthews DG, Kawamoto C, Harris CJ, Quinn JF, Soumyanath A. Centella asiatica attenuates Aβ-induced neurodegenerative spine loss and dendritic simplification. Neurosci Lett 2017; 646: 24-29
    CrossrefPubMedGoogle Scholar
  • 12 Perron JC, Bixby JL. Distinct neurite outgrowth signaling pathways converge on ERK activation. Mol Cell Neurosci 1999; 13: 362-378
    CrossrefPubMedGoogle Scholar
  • 13 Read D, Gorman A. Involvement of Akt in neurite outgrowth. Cell Mol Life Sci 2009; 66: 2975-2984
    CrossrefPubMedGoogle Scholar
  • 14 Xu Y, Cao Z, Khan I, Luo Y. Gotu Kola (Centella asiatica) extract enhances phosphorylation of cyclic AMP response element binding protein in neuroblastoma cells expressing amyloid beta peptide. J Alzheimers Dis 2008; 13: 341-349
    CrossrefPubMedGoogle Scholar
  • 15 Lin J, Jiang H, Ding X. Synergistic combinations of five single drugs from Centella asiatica for neuronal differentiation. Neuroreport 2017; 1: 23-27
    PubMedGoogle Scholar
  • 16 Harrill JA, Mundy WR. Quantitative assessment of neurite outgrowth in PC12 cells. Methods Mol Biol 2011; 758: 331-348
    PubMedGoogle Scholar
  • 17 Kumar V, Zhang MX, Swank MW, Kunz J, Wu GY. Regulation of dendritic morphogenesis by Ras-PI3K-Akt-mTOR and Ras-MAPK signaling pathways. J Neurosci 2005; 25: 11288-11299
    CrossrefPubMedGoogle Scholar
  • 18 Lin X, Huang R, Zhang S, Wei L, Zhuo L, Wu X, Tang A, Huang Q. Beneficial effects of asiaticoside on cognitive deficits in senescence-accelerated mice. Fitoterapia 2013; 87: 69-77
    CrossrefPubMedGoogle Scholar
  • 19 da Silva JS, Dotti CG. Breaking the neuronal sphere: regulation of the actin cytoskeleton in neuritogenesis. Nat Rev Neurosci 2002; 9: 694-704
    PubMedGoogle Scholar
  • 20 Lonze BE, Ginty DD. Function and regulation of CREB family transcription factors in the nervous system. Neuron 2002; 4: 605-623
    PubMedGoogle Scholar
  • 21 Xiao J, Liu Y. Differential roles of ERK and JNK in early and late stages of neuritogenesis: a study in a novel PC12 model system. J Neurochem 2003; 6: 1516-1523
    PubMedGoogle Scholar
  • 22 Ebisuya M, Kondoh K, Nishida E. The duration, magnitude and compartmentalization of ERK MAP kinase activity: mechanisms for providing signaling specificity. J Cell Sci 2005; 118: 2997-3002
    CrossrefPubMedGoogle Scholar
  • 23 von Kriegsheim A, Baiocchi D, Birtwistle M, Sumpton D, Bienvenut W, Morrice N, Yamada K, Lamond A, Kalna G, Orton R, Gilbert D, Kolch W. Cell fate decisions are specified by the dynamic ERK interactome. Nat Cell Biol 2009; 11: 1458-1464
    CrossrefPubMedGoogle Scholar
  • 24 Kao S, Jaiswal RK, Kolch W, Landreth GE. Identification of the mechanisms regulating the differential activation of the MAPK cascade by epidermal growth factor and nerve growth factor in PC12 cells. J Biol Chem 2001; 21: 18169-18177
    PubMedGoogle Scholar
  • 25 Schoeberl B, Eichler-Jonsson C, Gilles ED, Müller G. Computational modeling of the dynamics of the MAP kinase cascade activated by surface and internalized EGF receptors. Nat Biotechnol 2002; 20: 370-375
    CrossrefPubMedGoogle Scholar
  • 26 Papadeas ST, Blake BL, Knapp DJ, Breese GR. Sustained extracellular signal-regulated kinase 1/2 phosphorylation in neonate 6-hydroxydopamine-lesioned rats after repeated D1-dopamine receptor agonist administration: implications for NMDA receptor involvement. J Neurosci 2004; 26: 5863-5876
    PubMedGoogle Scholar
  • 27 Ha S, Redmond L. ERK mediates activity dependent neuronal complexity via sustained activity and CREB-mediated signaling. Dev Neurobiol 2008; 68: 1565-1579
    CrossrefPubMedGoogle Scholar
  • 28 Hur J, Lee P, Moon E, Kang I, Kim SH, Oh MS, Kim SY. Neurite outgrowth induced by spicatoside A, a steroidal saponin, via the tyrosine kinase A receptor pathway. Eur J Pharmacol 2009; 620: 9-15
    CrossrefPubMedGoogle Scholar
  • 29 Kim MS, Yu JM, Kim HJ, Kim HB, Kim ST, Jang SK, Choi YW, Lee DI, Joo SS. Ginsenoside Re and Rd enhance the expression of cholinergic markers and neuronal differentiation in Neuro-2a cells. Biol Pharm Bull 2014; 37: 826-833
    CrossrefPubMedGoogle Scholar
  • 30 Zhou H, Xue W, Chu SF, Wang ZZ, Li CJ, Jiang YN, Luo LM, Luo P, Li G, Zhang DM, Chen NH. Polygalasaponin XXXII, a triterpenoid saponin from Polygalae Radix, attenuates scopolamine-induced cognitive impairments in mice. Acta Pharmacol Sin 2016; 37: 1045-1053
    CrossrefPubMedGoogle Scholar
  • 31 Gu X, Meng S, Liu S, Jia C, Fang Y, Li S, Fu C, Song Q, Lin L, Wang X. miR-124 represses ROCK1 expression to promote neurite elongation through activation of the PI3K/Akt signal pathway. J Mol Neurosci 2014; 1: 156-165
    PubMedGoogle Scholar
  • 32 Hur EM, Zhou FQ. GSK3 signalling in neural development. Nat Rev Neurosci 2010; 11: 539-551
    PubMedGoogle Scholar
  • 33 Mullen LM, Pak KK, Chavez E, Kondo K, Brand Y, Ryan AF. Ras/p38 and PI3K/Akt but not Mek/Erk signaling mediate BDNF-induced neurite formation on neonatal cochlear spiral ganglion explants. Brain Res 2012; 1430: 25-34
    CrossrefPubMedGoogle Scholar
  • 34 Liu J, Shimizu K, Tanaka A, Shinobu W, Ohnuki K, Nakamura T, Kondo R. Target proteins of ganoderic acid DM provides clues to various pharmacological mechanisms. Sci Rep 2012; 2: 905
    CrossrefPubMedGoogle Scholar
  • 35 Chen YC, Liu YL, Li FY, Chang CI, Wang SY, Lee KY, Li SL, Chen YP, Jinn TR, Tzen JT. Antcin A, a steroid-like compound from Antrodia camphorata, exerts anti-inflammatory effect via mimicking glucocorticoids. Acta Pharmacol Sin 2011; 32: 904-911
    CrossrefPubMedGoogle Scholar
  • 36 Honda T, Rounds BV, Bore L, Finlay HJ, Favaloro jr. FG, Suh N, Wang Y, Sporn MB, Gribble GW. Synthetic oleanane and ursane triterpenoids with modified rings A and C: a series of highly active inhibitors of nitric oxide production in mouse macrophages. J Med Chem 2000; 43: 4233-4246
    CrossrefPubMedGoogle Scholar
  • 37 Rafat M, Fong KW, Goldsipe A, Stephenson BC, Coradetti ST, Sambandan TG, Sinskey AJ, Rha C. Association (micellization) and partitioning of aglycon triterpenoids. J Colloid Interface Sci 2008; 2: 324-330
    PubMedGoogle Scholar
  • 38 Dong MS, Jung SH, Kim HJ, Kim JR, Zhao LX, Lee ES, Lee EJ, Yi JB, Lee N, Cho YB, Kwak WJ, Park YI. Structure-related cytotoxicity and anti-hepatofibric effect of asiatic acid derivatives in rat hepatic stellate cell-line, HSC-T6. Arch Pharm Res 2004; 27: 512-517
    CrossrefPubMedGoogle Scholar
  • 39 Zhang Z, Li X, Li D, Luo M, Li Y, Song L, Jiang X. Asiaticoside ameliorates beta-amyloid-induced learning and memory deficits in rats by inhibiting mitochondrial apoptosis and reducing inflammatory factors. Exp Ther Med 2017; 13: 413-420
    CrossrefPubMedGoogle Scholar
  • 40 Welbat JU, Sirichoat A, Chaijaroonkhanarak W, Prachaney P, Pannangrong W, Pakdeechote P, Sripanidkulchai B, Wigmore P. Asiatic acid prevents the deleterious effects of valproic acid on cognition and hippocampal cell proliferation and survival. Nutrients 2016; 8: 1-11
    CrossrefPubMedGoogle Scholar
  • 41 Yuan Y, Zhang H, Sun F, Sun S, Zhu Z, Chai Y. Biopharmaceutical and pharmacokinetic characterization of asiatic acid in Centella asiatica as determined by a sensitive and robust HPLC-MS method. J Ethnopharmacol 2015; 163: 31-38
    CrossrefPubMedGoogle Scholar
  • 42 Hengjumrut P, Anukunwithaya T, Tantisira MH, Tantisira B, Khemawoot P. Comparative pharmacokinetics between madecassoside and asiaticoside presented in a standardised extract of Centella asiatica, ECa 233 and their respective pure compound given separately in rats. Xenobiotica 2018; 48: 18-27
    CrossrefPubMedGoogle Scholar
  • 43 Anukunwithaya T, Tantisira MH, Tantisira B, Khemawoot P. Pharmacokinetics of a standardized extract of Centella asiatica ECa 233 in rats. Planta Med 2016; 83: 710-717
    Article in Thieme ConnectPubMedGoogle Scholar