Sains Malaysiana 47(11)(2018): 2783–2788

http://dx.doi.org/10.17576/jsm-2018-4711-21

 

The Effect of Centella asiatica (L.) Urban on the Organotypic Model of Spinal Cord Injury

(Kesan Centella asiatica (L.) Urban pada Model Organotip Kecederaan Saraf Tunjang)

 

NUR NABILAH AHMAD PUZI1,2,3, YOGESWARAN LOKANATHAN1 & RUSZYMAH BT HJ IDRUS1,2*

 

1Tissue Engineering Centre, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaacob Latif, Bandar Tun Razak, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

 

2Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre

Jalan Yaacob Latif, Bandar Tun Razak, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

 

3Center for Craniofacial Diagnostics and Biosciences, Faculty of Dentistry, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Federal Territory, Malaysia

 

Received: 28 March 2018/Accepted: 30 July 2018

 

ABSTRACT

Centella asiatica (L.) Urban (CA) is a well- known plant used to improve brain and memory functions in traditional medicine. Scientifically it was proven to show neurogenic effect on neural cell lines and in rat’s hippocampus. Its effect on spinal cord (SC) neurons, however, have not been studied. Aim of this study was to investigate the effects of raw extract of CA (RECA) on neurite outgrowths in an organotypic model of SC injury (OMSCI). OMSCI was prepared using SC slices obtained from postnatal-day 8 rat pups. Spinal cord tissues were embedded in gelatine gel and sliced to produce 300 μm thick slices. These slices were 100% viable for 8 days in culture. RECA, in concentrations of 0-800 μg/mL was added to the OMSCI media for 7 days, followed by immunostaining for TUJ-1 and GFAP. The investigated parameters were mean neurite count, mean neurite length, mean longest neurite and growth ratio. The tested RECA concentrations showed no cytotoxicity. ANOVA and Kruskal-Wallis tests showed no significant difference between groups in all the tested parameters. This may be due to low content of neurotrophic bioactive compounds content in the extract, which probably due to differences in geographical location, extraction method and absence of neurotrophic factors in the media. In conclusion, the tested RECA concentration were found to be safe; but without notable neurotrophic effects on the spinal cord organotypic model as demonstrated in this study.

 

Keywords: Centella asiatica; interface culture; neurite measurements; neurotrophin

 

ABSTRAK

Centella asiatica (L.) Urban (CA) adalah tumbuhan terkenal yang digunakan untuk meningkatkan fungsi otak dan ingatan dalam perubatan tradisi. Secara saintifik, ia terbukti menunjukkan kesan neurogenik pada sel-sel sel saraf dan hipokampus tikus. Kesannya pada neuron saraf tunjang (ST), bagaimanapun, belum dikaji. Tujuan penyelidikan ini adalah untuk mengkaji kesan ekstrak mentah CA (RECA) ke atas neurit dalam model organotip kecederaan ST (MOKST). MOKST telah disediakan menggunakan kepingan ST yang diperoleh daripada tikus berumur 8 hari posnatal. Tisu saraf tunjang dibenamkan dalam gel gelatin dan dihiris untuk menghasilkan kepingan berketebalan 300 μm. Kepingan ini menunjukkan 100% kebolehhidupan apabila dikultur selama 8 hari. RECA, dengan kepekatan 0-800 μg/mL telah ditambahkan kepada media MOKST selama 7 hari, diikuti dengan pewarnaan imunologi untuk TUJ-1 dan GFAP. Parameter yang dikaji adalah purata bilangan neurit, panjang neurit, neurit terpanjang dan nisbah pertumbuhan. Kepekatan RECA yang diuji tidak menunjukkan kesitotoksikan. Ujian ANOVA dan Kruskal-Wallis tidak menunjukkan perbezaan yang signifikan antara kumpulan bagi semua parameter yang diuji. Ini mungkin disebabkan oleh kandungan bioaktif neurotrof yang rendah dalam ekstrak, yang mungkin disebabkan oleh perbezaan lokasi geografi, kaedah pengekstrakan dan ketiadaan faktor neurotrof dalam media. Kepekatan RECA yang diuji didapati selamat; namun tidak menunjukkan kesan neurotrof pada model organotip saraf tunjang seperti yang diperhatikan dalam kajian ini.

 

Kata kunci: Centella asiatica; kultur antara-muka; neurotrofin; pengukuran neurit

REFERENCES

Alqahtani, A., Tongkao-on, W., Li, KM, Razmovski-Naumovski, V., Chan, K. & Li, G.Q. 2015. Seasonal variation of triterpenes and phenolic compounds in Australian Centella asiatica (L.) Urb. Phytochemical Analysis 26: 436-443.

Azwanida, N.N. 2015. A review on the extraction methods use in medicinal plants, principle, strength and limitation. Medicinal & Aromatic Plants 4: 3-8.

Bhavna, D. & Jyoti, K. 2011. Centella asiatica: The elixir of life. International Journal of Research in Ayurveda & Pharmacy 2: 431-438.

Chew, K.K., Ng, S.Y., Thoo, Y.Y., Khoo, M.Z., Wan Aida, W.M. & Ho, C.W. 2011. Effect of ethanol concentration, extraction time and extraction temperature on the recovery of phenolic compounds and antioxidant capacity of orthosiphon stamineus extracts. International Food Research Journal 18: 1427-1435.

De Simoni, A. & Yu, L.M.Y. 2006. Preparation of organotypic hippocampal slice cultures: Interface method. Nature Protocols 1(3): 1439-1445.

Gähwiler, B.H., Capogna, M., Debanne, D., McKinney, R.A. & Thompson, S.M. 1997. Organotypic slice sultures: A technique has come of age. Trends Neuroscience 20: 471-477.

Gurcan, O., Gurcay, A.G., Kazanci, A., Senturk, S., Bodur, E., Karaca, E.U., Turkoglu, O.F. & Bavbek, M. 2015. Effect of asiatic acid on the treatment of spinal cord injury: An experimental study in rats. Turkish Neurosurgery. doi: 10.5137/1019-5149.jtn.15747-15.2.

Huang, E.J. & Reichardt, L.F. 2001. Neurotrophins: Roles in neuronal development and function. Annual Review of Neuroscience 24: 677-736.

Ibrahim, A., Lee, K.Y., Kanoo, L.L., Tan, C.H., Hamid, M.A., Hamedon, M.A. & Haniff, J. 2013. Epidemiology of spinal cord injury in Hospital Kuala Lumpur. Spine 38: 419-424.

Jiang, W., Li, M., He, F., Bian, Z., He, Q., Wang, X., Yao, W. & Zhu, L. 2016. Neuroprotective effect of asiatic acid against spinal cord injury in rats. Life Sciences 157: 45-51.

Lokanathan, Y., Omar, N., Ahmad Puzi, N.N., Saim, A. & Hj Idrus, R. 2016. Recent updates in neuroprotective and neuroregenerative potential of Centella asiatica. Malaysian Journal of Medical Sciences 23: 4-14.

Morrison, B., Saatman, K.E., Meaney, D.F. & Mcintosh, T.K. 1998. In vitro central nervous system models of mechanically induced trauma: A Review. Journal of Neurotrauma 15(11): 911-928.

Norenberg, M.D., Smith, J. & Marcillo, A. 2004. The pathology of human spinal cord injury: Defining the problems. Journal of Neurotrauma 21: 429-440.

Nurlaily, A., Noor Baitee, A.R. & Musalmah, M. 2012. Comparative antioxidant and anti-inflammatory activity of different extracts of Centella asiatica (L.) Urban and its active compounds, asiaticoside and madecassoside. Med. & Health 7(2): 62-72.

Omar, N., Lokanathan, Y. & Hj. Idrus, R. 2017. Comparison of cytotoxicity measurements of Centella asiatica (L.) on Human Wharton’s Jelly-derived mesenchymal stem cells in vitro via mtt and presto blue assay 5: 10-18.

Othman, F. 2003. Chemical constituent & biological activities of Flavonoid from hydroponically grown pegaga. Thesis. Universiti Putra Malaysia (Unpublished).

Puttarak, P. & Panichayupakaranant, P. 2012. Factors affecting the content of pentacyclic triterpenes in Centella asiatica raw materials. Pharmaceutical Biology 50: 1508-1512.

Ramer, L.M., Ramer, M.S. & Bradbury, E.J. 2014. Restoring function after spinal cord injury: Towards clinical translation of experimental strategies. The Lancet Neurology 13: 1241- 1256.

Rao, M., Gadahad, K., Rao, M. & Rao, G. 2008. Enhancement of hippocampal CA3 neuronal dendritic arborization by Centella asiatica (Linn) fresh leaf extract treatment in adult rats. Journal of the Chinese Medical Association 71: 6-13.

Sadowsky, C., Volshteyn, O., Schultz, L. & McDonald, J.W. 2002. Spinal cord injury. Disability and Rehabilitation 24: 680-687.

Shamir, E.R. & Ewald, A.J. 2014. Three-dimensional organotypic culture: Experimental models of mammalian biology and disease. Nature Reviews Molecular Cell Biology 15(10): 647-664.

Shinomol, G.K., Muralidhara, M.S. & Bharath, M. 2011. Exploring the role of ‘Brahmi’ (Bocopa monnieri and Centella asiatica) in brain function and therapy. Recent Patents on Endocrine, Metabolic & Immune Drug Discovery 5: 33-49.

Silva, N.A., Sousa, N., Reis, R.L. & Salgado, A.J. 2014. From basics to clinical: A comprehensive review on spinal cord injury. Progress in Neurobiology 114: 25-57.

Soumyanath, A., Zhong, Y.P., Gold, S.A., Yu, X., Koop, D.R., Bourdette, D. & Gold, B.G. 2005. Centella asiatica accelerates nerve regeneration upon oral administration and contains multiple active fractions increasing neurite elongation in-vitro. The Journal of Pharmacy and Pharmacology 57: 1221-1229.

Stoppini, L., Buchs, P.A. & Muller, D. 1991. A simple method for organotypic cultures of nervous tissue. Journal of Neuroscience Methods 37(2): 173-182.

Sypecka, J., Koniusz, S., Kawalec, M. & Sarnowska, A. 2015. The organotypic longitudinal spinal cord slice culture for stem cell study. Stem Cells International 2015: Article ID. 471216.

Wanakhachornkrai, O., Pongrakhananon, V., Chunhacha, P., Wanasuntronwong, A., Vattanajun, A., Tantisira, B., Chanvorachote, P. & Tantisira, M.H. 2013. Neuritogenic effect of standardized extract of Centella asiatica ECa233 on human neuroblastoma cells. BMC Complementary and Alternative Medicine 13: 204.

Yang, Z. & Xiong, H.R. 2012. In vitro tissue-based models as a replacement for animal models in testing of drugs at the preclinical stages. In Biomedical Tissue Culture, edited by Ceccherini-Nelli, L. & Matteoli, B. InTechOpen. pp. 73-80.

Yung, Ong Hooi, Mohamad Yusof Maskat & Wan Aida Wan Mustapha. 2010. Effect of extraction on polyphenol content, antioxidant activity and pH in pegaga (Centella asiatica) extract. Sains Malaysiana 39(5): 747-752.

Zhang, X.G., Han, T., Zhang, Q.Y., Zhang, H., Huang, B.K., Xu, L.L. & Qin, L.P. 2009. Chemical fingerprinting and hierarchical clustering analysis of Centella asiatica from different locations in China. Chromatographia 69: 51-57.

 

 

*Corresponding author; email: ruszyidrus@gmail.com

 

 

 

 

 

 

 

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