Centella asiatica (Gotu kola) ethanol extract up-regulates hippocampal brain-derived neurotrophic factor (BDNF), tyrosine kinase B (TrkB) and extracellular signal-regulated protein kinase 1/2 (ERK1/2) signaling in chronic electrical stress model in rats

Document Type: Short Communication


1 Department of Anatomy, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia

2 Department of Surgery, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia

3 Division of Cardiovascular Medicine, Department of Internal Medicine, Kobe University Graduate School of Medicine, Kobe, Japan

4 Laboratory of Clinical Pharmaceutical Science, Kobe Pharmaceutical University, Kobe, Japan


Objective(s): Impairment of hippocampus function as a center for memory processing occurs due to stress. Centella asiatica L. (Gotu kola) is known to improve memory, intelligence, and neural protection although the precise mechanism is not well understood. This study aimed to investigate the effects of ethanol extracts of C. asiatica toward MAPK expression as down-stream signaling of brain-derived neurotrophic factor (BDNF).
Materials and Methods: We performed a chronic electrical stress model on 20 male Sprague Dawley rats (2 months-old, 180–200 g). Rats were divided into four groups: normal control group (Control) which received distilled water, and three treatment groups receiving oral Gotu kola ethanol extracts in oral doses of 150 mg/kg BW (CeA150), 300 mg/kg BW (CeA300), and 600 mg/kg BW (CeA600) over four weeks. Memory acquisition was assessed with Morris water maze. Hippocampus was harvested, then extracted for protein and RNA analysis. MAPK proteins (p38, ERK1/2, JNK) were measured using Western blot, meanwhile, BDNF and TrkB receptor were analyzed with real-time PCR (RT-PCR).
Results: CeA600 group revealed improvement of memory performance as shown by reduction in time and distance parameters compared to control during escape latency test. This finding associated with significant elevation of hippocampal BDNF protein and mRNA level with up-regulation of TrkB mRNA expression in CeA600 group compared to control. Western-blot analysis showed significant up-regulation of ERK1/2 protein level in CeA600 group (P<0.05) compare to control.
Conclusion: BDNF signaling through TrkB and ERK1/2 pathway contributes significantly to amelioration of memory performance after C. asiatica treatment in electrical stress model.


Main Subjects

1.    Mohamed S, Baqutayan S. Stress and coping mechanisms : a historical overview. Mediterr J Soc Sci. 2015;6(2):479–88.

2.    Esch T, Fricchione GL, Hospital MG. The role of stress in neurodegenerative diseases. Neuroendocrinol Lett. 2002;23(July):199–208.

3.    Mcewen BS, Nasca C, Gray JD. Stress effects on neuronal structure : hippocampus , amygdala , and prefrontal cortex. Neuropsychopharmacology. 2015;41(1):3–23.

4.    Kim EJ, Pellman B, Kim JJ. Stress effects on the hippocampus : a critical review. Learn Mem. 2015;22:411–6.

5.    Yamada K, Mizuno M NT. Role for brain-derived neurotrophic factor in learning and memory. Life Sci. 2002;70(7):735–44.

6.    Mizui T, Kojima M. Recent Advances in the biology of BDNF and the newly identified. J Neurol Neuromedicine. 2018;3(6):1–4.

7.    Cunha C, Brambilla R, Thomas KL. A simple role for BDNF in learning and memory ? Mol Neurosci. 2010;3(February):1–14.

8.    Mizuno M, Yamada K, Olariu A, Nawa H, Nabeshima T. Involvement of Brain-Derived Neurotrophic Factor in spatial memory formation and maintenance in a radial arm maze test in rats. J Neurosci. 2000;20(18):7116–21.

9.    Grønli J, Bramham C, Murison R, Kanhema T, Fiske E. Chronic mild stress inhibits BDNF protein expression and CREB activation in the dentate gyrus but not in the hippocampus proper. Pharmacol Biochem Behav. 2007;85(2006):842–9.

10. Shi S, Shao S, Yuan B, Pan F, Li Z. Acute stress and chronic stress change Brain-Derived Neurotrophic Factor (BDNF) and tyrosine kinase-coupled receptor (TrkB) expression in both young and aged rat hippocampus. Yonsei Med J. 2010;51(5):661–71.

11. Qi F, Yang L, Tian Z, Zhao M, Liu S, An J. Neuroprotective effects of asiaticoside. Neural Regen Res. 2014;9(13):1275–82.

12. Soumyanath A, Zhong Y-P, Yu X, Bourdette D, Koop DR, Gold SA, et al. Centella asiatica accelerates nerve regeneration upon oral administration and contains multiple active fractions increasing neurite elongation in-vitro. J Pharm Pharmacol. 2005;57(9):1221–9.

13. Sari DCR, Aswin S, Susilowati R, Ar-rochmah M, Prakosa D, Tranggono U, et al. Ethanol extracts of Centella asiatica leaf improves memory performance in rats after chronic stress via reducing nitric oxide and increasing Brain-Derived Neurotrophic Factor (BDNF) concentration. Int J Psychol. 2014;1(1):61–7.

14. Mizuno M, Yamada K, Takei N, Tran MH, He J, Nakajima A, et al. Phosphatidylinositol 3-kinase : a molecule mediating BDNF-dependent spatial memory formation. Mol Pychiatry. 2003;8:217–24.

15. Alonso M, Medina JH, Pozzo-miller L. ERK1/2 activation is necessary for BDNF to increase dendritic spine density in hippocampal CA1 pyramidal neurons. Learn Mem. 2004;11:172–8.

16. Sari DCR. Efek neurotrofik dan neuroprotektif ekstrak ethanol daun pegagan (Centella asiatica (L.) Urb.) terhadap gangguan memori spasial pasca-stres kronik. Gadjah Mada; 2015.

17. Liu W, Ge T, Leng Y, Pan Z, Fan J, Yang W, et al. The role of neural plasticity in depression : from hippocampus to prefrontal cortex. Hindawi Neural Plasicity. 2017;2017:1–11.

18. Boondam Y, Songvut P, Tantisira MH, Ta S. Inverted U-shaped response of a standardized extract of Centella asiatica (ECa 233) on memory enhancement. Nat Sci Reports. 2019;9(8404):1–11.

19. Yamada K, Nabeshima T. Current perspective Brain-Derived Neurotrophic Factor/TrkB signaling in memory processes. J Phamracol Sci. 2003;270:267–70.

20. Hacioglu G, Senturk A, Ince I, Alver A. Assessment of oxidative stress parameters of Brain‐Derived Neurotrophic Factor heterozygous mice in acute stress model. Iran J Basic Med Sci. 2015;19(4):388–93.

21. Yun K, Kim J, Kim J, Lee K, Jeong S, Park H, et al. Inhibition of LPS-induced NO and PGE 2 production by asiatic acid via NF-κ B inactivation in RAW 264 . 7 macrophages : possible involvement of the IKK and MAPK pathways. Int Immunopharmacol. 2008;8:431–41.

22. Orhan IE. Centella asiatica (L.) Urban: from traditional medicine to modern medicine with neuroprotective potential. Evidence-based Complement Altern Med. 2012;2012:1–9.

23. Musfiroh IDA, Muhtadi A, Kartasasmita RE, Tjahjono DH. In silico study of asiatic acid interaction with inducible nitric oxide synthase (iNOS) and cyclooygenase-2 (COX-2). Int J Pharm Pharm Sci. 2013;5:204–7.

24. Barbieri SS, Eligini S, Brambilla M, Tremoli E, Colli S. Reactive oxygen species mediate cyclooxygenase-2 induction during monocyte to macrophage differentiation : critical role of NADPH oxidase. Cardiovasc Res. 2003;60:187–97.

25. Lin W, Peng Y, Hou C. Ferulic acid protects PC12 neurons against hypoxia by inhibiting the p-MAPKs and COX-2 pathways. Iran J Basic Med Sci. 2014;18(5):478–84.

26. Gray NE, Zweig JA, Caruso M, Martin MD, Zhu JY, Quinn JF, et al. Centella asiatica increases hippocampal synaptic density and improves memory and executive function in aged mice. Brain Behav. 2018;8(e01024):1–11.

27. Veerendra Kumar M., Gupta Y. Effect of different extracts of Centella asiatica on cognition and markers of oxidative stress in rats. J Ethnopharmacol. 2002;79(2):253–60.

28. Veerendra Kumar M, Gupta Y. Effect of Centella asiatica on cognition and oxidative stress in an intracerebroventricular streptozotocin model of Alzheimer’s disease in rats. Clin Exp Pharmacol Physiol. 2003;30(5–6).

29. Xu M, Xiong Y, Liu J, Qian J, Zhu L, Gao J. Asiatic acid, a pentacyclic triterpene in Centella asiatica, attenuates glutamate-induced cognitive deficits in mice and apoptosis in SH-SY5Y cells. Acta Pharmacol Sin. 2012;33(5):578–87.

30. Xu C, Qu R, Zhang J, Li L, Ma S. Neuroprotective effects of madecassoside in early stage of Parkinson’s disease induced by MPTP in rats. Fitoterapia. 2013;90:112–8.

31. Ramanathan M, Sivakumar S, Anandvijayakumar P, Saravanababu C, Pandian P. Neuroprotective evaluation of standardized extract of Centella asiatica in monosodium glutamate treated rats. Indian J Exp Biol. 2007;45(5):425–31.

32. Subathra M, Shila S, Devi MA, Panneerselvam C. Emerging role of Centella asiatica in improving age-related neurological antioxidant status. Exp Gerontol. 2005;40(8–9):707–15.

33. Rao SB, Chetana M, Uma Devi P. Centella asiatica treatment during postnatal period enhances learning and memory in mice. Physiol Behav. 2005;86(4):449–57.

34. Nagappan G, Lu B. Activity-dependent modulation of the BDNF receptor TrkB : mechanisms and implications. Trends Neurosci. 2005;28(9):464–71.

35. Bozon B, Kelly Á, Josselyn SA, Silva AJ, Davis S, Laroche S, et al. MAPK , CREB and zif268 are all required for the consolidation of recognition memory. J Biol Sci. 2003;358:805-14

36. Kelly A, Laroche S, Sabrina D. Activation of Mitogen-Activated Protein Kinase/Extracellular Signal-Regulated Kinase in hippocampal circuitry is required for consolidation and reconsolidation of recognition memory. J Neurosci. 2003;12(23):5354–60.

37. Lonze BE, Ginty DD. Function and regulation of CREB family transcription factors in the nervous system CREB and its close relatives are now widely accepted. Neuron. 2002;35:605–23.

38. Ortega-martínez S. A new perspective on the role of the CREB family of transcription factors in memory consolidation via adult hippocampal neurogenesis. Mol Neurosci. 2015;8(August):1–12.

39. Toyoda H, Zhao M, Xu H, Wu L, Ren M, Zhuo M. Requirement of Extracellular Signal-Regulated Kinase/Mitogen-Activated Protein Kinase for long-term potentiation in adult mouse anterior cingulate cortex. Mol Pain. 2007;3(36):1–15.

40. Schafe GE, Ledoux JE. Memory consolidation of auditory pavlovian fear conditioning requires protein synthesis and protein kinase a in the amygdala. J Neurosci. 2000;20:1–5.

41. Ying S, Futter M, Rosenblum K, Webber MJ, Hunt SP, Bliss TVP, et al. Brain-Derived Neurotrophic Factor induces long-term potentiation in intact adult hippocampus : requirement for ERK activation coupled to CREB and upregulation of Arc synthesis. J Neurosci. 2002;22(5):1532–40.

42. Lee S, Yang M, Kim J, Son Y, Kim J, Ahn W, et al. Involvement of BDNF/ERK signaling in spontaneous recovery from trimethyltin-induced hippocampal neurotoxicity in mice. Brain Res Bull. 2016;121:48–58.

43. Silva-peña D, Rivera P, Alén F, Vargas A, Rubio L, García-marchena N, et al. Oleoylethanolamide modulates BDNF-ERK signaling and neurogenesis in the hippocampi of rats exposed to 9-THC and ethanol binge drinking during adolescence. Mol Neurosci. 2019;12(April):1–15.