The effect of Centella asiatica L. Urban. and Curcuma longa L. extracts combination in improving memory performance in stroke model rats and its acute toxicity

  • Abdul Gofir Department of Neurology, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada/Dr. Sardjito General Hospital
  • Mawaddah Ar Rochmah Department of Neurology, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada/Dr. Sardjito General Hospital
  • Samekto Wibowo Department of Neurology, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada/Dr. Sardjito General Hospital
  • Mohammad Hakimi Department of Obstetrics and Gynecology, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada/Dr. Sardjito General Hospital
  • Mustofa Department of Pharmacology and Therapy, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia
Keywords: Centella asiatica L. Urban, Curcuma longa L., stroke, cognitive impairment, memory


Post-stroke cognitive impairment involves memory, visuoconstructional, and

spatial dysfunctions. Centella asiatica L. Urban. and Curcuma longa L. are both

well-known herbs in South and South-East Asia countries that are believed to

possess neuroprotective properties as memory enhancers. This study aimed to

investigate the effects of C. asiatica L. Urban. and C. longa L. extracts combination

in improving memory performance in stroke model rats and its acute toxicity.

Twenty-five Wistar rats underwent transient bilateral common carotid artery

occlusion. Y-maze pre-test was performed 24 h after the occlusion. The rats were

then divided into five groups i.e. Group 1 received NaCMC dilution, Group 2

received donepezil 0. 7 mg/kg BW/day, and Groups 3, 4, and 5 received the extracts

combination with dose of 59; 118; and 236 mg/kg BW/day, respectively. Y-maze

post-test was performed 24 h after the last dose had been given. Spontaneous

alternation behavior was used as the indicator of working memory measurement.

The fixed-dose method according to OECD Guideline was used to estimate the

LD50 in acute toxicity testing on Wistar rats. There was a significant difference

in the delta spontaneous alternation percentage among groups tested (p<0. 05).

Group 1 had significant difference with any other group (compared to Groups

2 to 5; p<0.05). No significant difference could be found between groups of the

extracts combination and donepezil group suggesting that the effect is not more

inferior nor dose-dependent in improving memory performance. The extracts

combination at a single dose of 2000 mg/kg BW did not show related signs of

toxicity or mortality in any of the rats during the 14-day observation period. In

conclusion, the C. asiatica L. Urban. and C. longa L. extracts combination can

improve performance of memory on stroke model animal. According to Globally

Harmonized Classification System, the extracts combination can be classified as

Category 5/unclassified.


Hostenbach J,Mulder T, van Limbeek J, Donders R, Schoonderwaldt H. Cognitive decline following stroke: a comprehensive study of cognitive decline following stroke. J Clin Exp Neurol 1998; 20: 503-17. https://doi. org/10. 1076/jcen. 20. 4. 503. 1471

Rasquin S, Lodder J, Verhey F. The association between psychiatric and cognitive symptoms after stroke: a prospective study. Cerebrovasc Dis 2005; 19(5): 309-16. https://doi. org/10. 1159/000084499

Ballard C, Stephens S, Kenny R, Kalaria R, Tovee M, O’Brien J. Profile of neuropsychological deficits in older stroke survivors without dementia. Dement Geriatr Cog Dis 2003; 16(1):52-6. https://doi. org/10. 1159/000069994

Tatemichi TK, Desmond DW, Stern Y, Paik M, Bagiella. Cognitive impairment after stroke: frequency, patterns, and relationship to functional abilities J Neurol Neurosurg Psychiatry 1994; 57:202-7. https://doi. org/10. 1136/jnnp. 57. 2. 202

Jaillard A, Naegele B, Trabucco- Miguel S, LeBas JF, Hommel M. Hidden dysfunctioning in sub acute stroke. Stroke 2009; 40: 2473-9. 541144

Nys GMS, van Zandvoort MJE, de Kort PLM, Jansen BPW, Den Haal EHF, Kappelle LJ, et al. Cognitive disorders in acute stroke: prevalence and clinical determination. Cerebrovasc Di 2007; 23(5-6):408-16.

Middleton LE, Lam B, Fahmi H, Black SE, Mcllroy WE, Stuss DT, et al. Frequency of domain-specific cognitive impairment in subacute and chronic stroke. Neuro Rehabilitation 2014; 34 (2): 305-12.

Mehla J, Gupta P, Pahuja M, Diwan D, Diksha D. Indian medicinal herbs and formulations for Alzheimer’s Disease, from traditional knowledge to scientific assessment. Brain Sci 2020; 101(2): 964. 3390brainsci10120964

Gohil KJ, Pattel JA, Gajjar AK. Pharmacological review on Centella asiatica : a potential herbal cure-all. Indian J Pharm Sci 2010: 72 (5):546-56. 78519

Sari DCR, Aswin S, Susilowati R, Ar- Rochmah M, Prakosa D, Romi M, 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. GSTF J of Psychol 2014; (1):1.

Farhana KM, Malueka RG, Wibowo S, Gofir A. Effectiveness of Gotu Kola extract 750 mg and 1000 mg compared with folic acid 3 mg in improving vascular cognitive impairment after stroke. Evid Based Complement Alternat Med 2016: 1-6. 1155/2016/2795915

Karting T. Clinical application of Centella asiatica (L) Urb. In: Herbs, spices, and medicinal plants (Cracker LE, Simon JE Eds.). Phoenix, AZ, USA: Oxyx Press. 1998: 145-73.

Chivapat S, Chavalittumrong P, Attawish A, Boonruad T, Bansiddhi J, Phadungpat S, et al. Toxicity study of Centella asiatica (L) urban. J Thai Trad Alternat Med 2004; 2: 3-17.

Perrone D, Ardito F, Giannatempo G, Dioguardi M, Troiano G, Lo Russo L, et al. Biological and therapeutic activities, and anticancer properties of curcumin. Expert Ther Med 2015;10(5):1615-23. https://doi. org/10. 3892/etm. 2015. 2749

Sharifi-Rad J, Rayess YE, Rizk AA, Sadaka C, Zgheib R, Zam W, et al . Turmeric and its major compound curcumin on health: bioactive effects and safety profiles for food, pharmaceutical, biotechnological and medicinal applications. Front Pharmacol 2020 15; 11: 01021. 01021

Xu Y, Lin D, Li S, Li G, Shyamala SG, Barish PA, et al. Curcumin reverses impaired cognition and neuronal plasticity induced by chronic stress. Neuropharmacology 2009; 57(4):463- 71. neuropharm.2009.06.010

Wang Y, Yin H, Li J, Zhang Y, Han B, Zeng Z, et al. Amelioration of β -amyloid-induced cognitive dysfunction and hippocampal axon degeneration by curcumin is associated with suppression of CRMP- 2 hyperphosphorylation. Neurosci Lett 2013; 557:112-7. neulet.2013.10.024

Yin HL, Wang YL, Li JF, Han B, Zhang XX, Wang YT, et al. Effects of curcumin on hippocampal expression of NgR and axonal regeneration in A β - induced cognitive disorder rats. Genet Mol Res 2014; 13(1):2039-47.

Baum L, Ng A. Curcumin interaction with copper and iron suggests one possible mechanism of action in Alzheimer’s Disease animal models, J Alzheimer Dis 2004; 6 (4):367-77.

McClure R, Ong H, Janve V, Barton S, Zhu M, Li B, et al. Aerosol delivery of curcumin reduced amyloid- deposition and improved cognitive performance in transgenic model of Alzheimer’s disease. J Alzheimer Dis 2016; 55 (2): 797-811. https://doi. org/10. 3233/JAD-160289

Kim SH & Lee HS. Acute oral toxicity study of ethanol extract of Curcuma longa L. in mice. J Life Sci 2014; 24(10): 1132-36

Handayani ES, Nurmasitoh T, Ahmad SA, Fauziah AN, Rizam R, Rahmawati RY, et al. Effect of BCCAO duration and animal models sex on brain ischemic volume after 24 hours reperfusion. Bangladesh J Med Sci 2018; 17(1): 129- 37.

Jokinen H, Melkas S, Yikoski R, Pohjasvaara T, Kaste M, Erkinjuntti, et al. Post-stroke cognitive impairment is common even after successful clinical recovery. Eur J Neurol, 2015; 22: 1288-94.

Kraeuter AK, Guest PC, and Sarnyai Z. The Y-Maze for assessment of spatial working and reference memory in mice. Methods Mol Biol 2019;1916:105-111. 8994-2_10

Sbrini G, Brivio P, Fumagalli M, Giavarini F, Caruso D, Racagni G, et al . Centella asiatica l. Phytosome improves cognitive performance by promoting bdnf expression in rat prefrontal cortex. Nutrients 2020; 12 (2): 355. https://doi. org/10. 3390/nu12020355

Sari DCR, Arfian N, Tranggono U, Setyaningsih WAW, Romi MM, Emoto N. 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. Iran J Basic Med Sci 2019; 22(10): 1218.

Wang R, Li YB, Li YH, Xu Y, Wu HL, Li XJ. Curcumin protects against glutamate excitotoxicity in rat cerebral cortical neurons by increasing brain-derived neurotrophic factor level and activating TrkB. Brain Res 2008; 1210: 84-91. 2008.01.104

Sbrini G, Brivio P, Sangiovanni E, Fumagalli M, Racagni G, Dell’Agli M, et al. Chronic treatment with a phytosomal preparation containing Centella asiatica L. and Curcuma longa L. affects local protein synthesis by modulating the BDNF- mTOR-S6 pathway. Biomedicines 2020; 8(12):544. biomedicines8120544

Palpu P, Rao CV, Kishore K, Gupta YK, Kartik R, Govindrajan R. U. S. Patent No. 7,429,397. Washington, DC: U. S. Patent and Trademark Office. 2008.

Astuti, Sutarni S, and Setyopranoto I. Serum brain-derived neurotrophic factor (BDNF) level may predict the functional outcome of acute ischemic stroke patients. Biomed Pharmacol J 2020; 13:4. https://doi. org/10. 13005/bpj/2075

Hassan TM, Yarube IU. Peripheral brain-derived neurotrophic factor is reduced in stroke survivors with cognitive impairment. Pathophysiology 2018; 25(4):405-10. pathophys.2018.08.003

Hsu CC, Kuo TW, Liu WP, Chang CP, Lin HJ. Calycosin preserves BDNF/ TrkB signaling and reduces post- stroke neurological injury after cerebral ischemia by reducing accumulation of hypertrophic and TNF- α -containing microglia in rats. J Neuroimmune Pharmacol 2020; 15 (2): 326-39. https://doi,org/10.1007/s11481-019- 09903-9.

Zhang ZH, Wu LN, Song JG, Li WQ. Correlations between cognitive impairment and brain derived neurotrophic factor expression in the hippocampus of post-stroke depression rats. Mol Med Rep 2012; 6(4): 889-93.

Luo, L, Li, C, Du, X, Shi, Q, Huang, Q, Xu X, et al. Effect of aerobic exercise on BDNF/proBDNF expression in the ischemic hippocampus and depression recovery of rats after stroke. Behav Brain Res 2019; 362: 323-31.