A NOVEL ANTI-PROLIFERATIVE ACTIVITY (EC50) OF PEGAGA (Centella asiatica) EXTRACT THROUGH IN VITRO 3-D CULTURE MICROENVIRONMENT
DOI:
https://doi.org/10.11113/jt.v79.8566Keywords:
Biomimetics, anti-proliferation, Centella asiatica, A549, IMR90, PHBV, 3-D scaffoldAbstract
Centella asiatica or pegaga is one of the botanical plants that consists of many phytochemicals and is known for being able to offer various effects on wound healing, as well as functioning as an antioxidant and anticancer property. Therefore, this study was carried out to determine the efficacy of Centella asiatica water and alcohol-based extracts on the anti-proliferative activity of human lung cancer cells (A549) and normal fibroblast (IMR90) by mean of in vitro 3-D cell culture system. A porous 3-D scaffold was fabricated from poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) using solvent-casting particulate-leaching (SCPL) process. Antioxidant analysis (total phenolic content (TPC), DPPH and FRAP assays) was conducted prior to the 3-D cell culture study. The results showed that the extract contained 7.50 ± 1.10 mg/ml of asiaticoside and 0.74 ± 0.24 mg/ml of madecassoside. These bio-active compounds were believed to inhibit the proliferation of cancer cells (A549). The availability of phenolic compounds in the extract (TPC: 10133 ± 119.30 mg/100 g) had proven that the antioxidant properties existed. Moreover, the other values obtained from the antioxidant analysis revealed its capacity as a good source of antioxidant (DPPH: 87 ± 1.0%; FRAP: 127 ± 14.98 mg/100 g). Next, the lung cancer cells (A549) were cultured using a two-dimensional (2-D) system to generate the IC50 value of 5.75 ± 1.0 µg/ml. The A549 cell viability (MTS assay) after a 3-day incubation exhibited a good sign of mortality for the both treated models ranging from 55% to 70% as compared to control one (without treatment) (p>0.05). However, when the extract was exposed to a normal fibroblast IMR90, the cell growth of the both treated models exhibited an almost 2-fold greater cell numbers than that of the untreated models (p<0.05) indicating that the extract did not possess any possible threat to a normal and healthy cell. Therefore, the use of Centella asiatica extracts in terminating cancer cells has been proven to be able to inhibit cell growth (greater than 40%) in just 3 days of incubation.Â
References
Yoo, H. H., Park, J. H. and Kwon, S. W. 2007. In vitro Cytotoxic Activity Of Some Korean Medicinal Plants On Human Cancer Cell Lines: Enhancement In Cytotoxicity By Heat Processing. Phytotherapy Research. 21(9): 900-903.
Singh, S., Gautam, A., Sharma, A. and Batra, A. 2010. Centella asiatica (L.): A Plant With Immense Medicinal Potential But Threatened. International Journal of Pharmaceutical Sciences Review and Research. 4(2).
Roopesh, C., Salomi, K. R., Nagarjuna, S. and Reddy, Y. P. 2011. Diuretic Activity Of Methanolic And Ethanolic Extracts Of Centella asiatica Leaves In Rats. International Research Journal of Pharmacy. 2(11): 163-165.
Coldren, C. D., Hashim, P., Ali, J. M., Oh, S.-K., Sinskey, A. J. and Rha, C. 2003. Gene Expression Changes In The Human Fibroblast Induced By Centella asiatica triterpenoids. Planta Medica. 69(8): 725-732.
Babu, T., Kuttan, G. and Padikkala, J. 1995. Cytotoxic And Anti-Tumour Properties Of Certain Taxa Of Umbelliferae With Special Reference To Centella asiatica (L.) Urban. Journal of Ethnopharmacalogy. 48(1): 53-57.
Gurfinkel, D. M., Chow, S., Hurren, R., Gronda, M., Henderson, C., Berube, C., Hedley, D. W. and Schimmer, A. D. 2006. Disruption Of The Endoplasmic Reticulum And Increases In Cytoplasmic Calcium Are Early Events In Cell Death Induced By The Natural Triterpenoid Asiatic Acid. Apoptosis. 11(9): 1463-1471.
Park, B. C., Bosire, K. O., Lee, E.-S., Lee, Y. S. and Kim, J.-A. 2005. Asiatic Acid Induces Apoptosis In SK-MEL-2 Human Melanoma Cells. Cancer Letters. 218(1): 81-90.
Lee, Y. S., Jin, D.-Q., Kwon, E. J., Park, S. H., Lee, E.-S., Jeong, T. C., Nam, D. H., Huh, K. and Kim, J.-A. 2002. Asiatic Acid, A Triterpene, Induces Apoptosis Through Intracellular Ca 2+ Release And Enhanced Expression Of p53 in HepG2 Human Hepatoma Cells. Cancer Letters. 186(1): 83-91
Tang, X.-L., Yang, X.-Y., Jung, H.-J., Kim, S.-Y., Jung, S.-Y., Choi, D.-Y., Park, W.-C. and Park, H. 2009. Asiatic Acid Induces Colon Cancer Cell Growth Inhibition And Apoptosis Through Mitochondrial Death Cascade. Biological and Pharmaceutical Bulletin. 32(8): 1399-1405.
Shimizu, T. 2014. Cell Sheet-Based Tissue Engineering For Fabricating 3-Dimensional Heart Tissues. Circ J. 78: 2594-2603.
Härmä, V., Virtanen, J., Mäkelä, R., Happonen, A., Mpindi, J.-P., Knuuttila, M., Kohonen, P., Lötjönen, J., Kallioniemi, O. and Nees, M. 2010. A Comprehensive Panel Of Three-Dimensional Models For Studies Of Prostate Cancer Growth, Invasion And Drug Responses. PLOS One. 5(5): e10431.
Babykutty, S., Padikkala, J., Sathiadevan, P., Vijayakurup, V., Azis, T., Srinivas, P. and Gopala, S. 2009. Apoptosis Induction Of Centella asiatica On Human Breast Cancer Cells. African Journal of Traditional, Complementary and Alternative Medicines. 6(1): 9-16.
Wong, S. P., Leong, L. P. and William Koh, J. H. 2006. Antioxidant Activities Of Aqueous Extracts Of Selected Plants. Food Chemistry 2006. 99(4): 775-783.
Brand-Williams, W., Cuvelier, M. and Berset, C. 1995. Use Of A Free Radical Method To Evaluate Antioxidant Activity. LWT-Food Science and Technology. 28(1): 25-30.
Zubairi, S. I., Bismarck, A., Koutinas, A., Panoskaltsis, N. and Mantalaris, A. 2013. Development Of Biomimetic PHB and PHBV Scaffolds For A Three Dimensional (3-D) In Vitro Human Leukaemia Model. Imperial College London.
Zubairi, S. I., Bismarck, A., Koutinas, A., Panoskaltsis, N. and Mantalaris, A. 2013. Surface Treatment of Poly (3-Hydroxybutyric Acid)(PHB) and Poly (3-Hydroxybutyric-co-3-Hydroxyvaleric Acid)(PHBV) Porous 3-D Scaffolds With An Improved Thickness To Enhance Cell-Biomaterial Adhesion and Interactions. Polymer. 1. 2: 3.
Zubairi, S. I., Koutinas, A., Bismarck, A., Panoskaltsis, N. and Mantalaris, A. 2011. Poly (3-hydroxybutyric acid)(PHB) and poly (3-hydroxybutyric-co-3-hydroxyvaleric acid)(PHBV) Porous Three-Dimensional Scaffolds With An Improved Thickness As Cell Growth Supporting Materials. Advances in Environmental Biology. 8(10): 925-932.
Aizad. S., Yahaya. B. H. and Zubairi S. I. 2014. Fabrication of 3-D Polymeric Scaffold: The Development of Continuous Flow Salt Leaching Kit Coupled with Ion Conductivity Detection. Regenerative Research. 3(2): 163-166.
Siripongvutikorn, S. 2012. Flavonoid, Phenolic Contents And Antioxidant Properties Of Thai Hot Curry Paste Extract And Its Ingredients As Affected Of Ph, Solvent Types And High Temperature.
Zheng, W. and Wang, S. Y. 2001. Antioxidant Activity And Phenolic Compounds In Selected Herbs. Journal of Agricultural And Food Chemistry. 49(11): 5165-5170.
James, J. T. and Dubery, I. A. 2009. Pentacyclic Triterpenoids From The Medicinal Herb, Centella asiatica (L.) Urban. Molecules. 14(10): 3922-3941.
Zainol, N., Voo, S., Sarmidi, M. R. and Azizi R. A. 2008. Profiling of Centella asiatica (L.) Urban Extract. The Malaysian Journal of Analytical Sciences. 12(2): 322-327.
Joshi, K. and Chaturvedi, P. 2013. Therapeutic Efficiency Of Centella asiatica (L.) urb. An Underutilized Green Leafy Vegetable: An Overview. International Journal of Pharma & Bio Sciences. 4(1): 135-149.
Jian, P., Guiqing, K., Chuanxun, Y., Beibei, Z., Risheng, J. and Yuan, Y. 2007. Separation And Determination Of Madecassic Acid In Extracts Of Centella asiatica Using High Performance Liquid Chromatography With β-Cyclodextrin As Mobile Phase Additive. Chinese Journal of Chromatography. 25(3): 316-318.
Gardner, R. S. 1969. The Use Of Tricine Buffer In Animal Tissue Cultures. The Journal Of Cell Biology. 42(1): 320-321.
Chew, Y. L., Chan, E. W. L., Tan, P. L., Lim, Y. Y., Stanslas, J. and Goh, J. K. 2011. Assessment Of Phytochemical Content, Polyphenolic Composition, Antioxidant And Antibacterial Activities Of Leguminosae Medicinal Plants In Peninsular Malaysia. BMC Complementary And Alternative Medicine. 11(1): 12.
Shukla, A., Rasik, A. M. and Dhawan, B. N. 1999. Asiaticosideâ€induced Elevation Of Antioxidant Levels In Healing Wounds. Phytotherapy Research. 13(1): 50-54.
Benzie, I. F. and Strain, J. 1996. The Ferric Reducing Ability Of Plasma (FRAP) As A Measure Of “Antioxidant Powerâ€: the FRAP Assay. Analytical Biochemistry. 239(1): 70-76.
Escarpa, A. and González, M. 2001. Approach To The Content Of Total Extractable Phenolic Compounds From Different Food Samples By Comparison Of Chromatographic And Spectrophotometric Methods. Analytica Chimica Acta. 427(1): 119-127.
Tulipani, S., Mezzetti, B., Capocasa, F., Bompadre, S., Beekwilder, J., De Vos, C. R., Capanoglu, E., Bovy, A. and Battino, M. 2008. Antioxidants, Phenolic Compounds, And Nutritional Quality Of Different Strawberry Genotypes. Journal of Agricultural and Food Chemistry. 56(3): 696-704;
Singleton, V. L., Orthofer, R. and Lamuela-Raventos, R. M. 1999. Analysis Of Total Phenols And Other Oxidation Substrates And Antioxidants By Means Of Folin-Ciocalteu Reagent. Methods in Enzymology. 299: 152-178.
Basile, A., Ferrara, L., Del Pezzo, M., Mele, G., Sorbo, S., Bassi, P. and Montesano, D. 2005. Antibacterial And Antioxidant Activities Of Ethanol Extract From Paullinia Cupana Mart. Journal of Ethnopharmacology. 102(1): 32-36.
Pittella, F., Dutra, R. C., Junior, D. D., Lopes, M. T. and Barbosa, N. R. 2009. Antioxidant And Cytotoxic Activities Of Centella asiatica (L) Urb. International Journal Of Molecular Sciences. 10(9): 3713-3721.
Buttke, T. M. and Sandstrom, P. A. 1994. Oxidative Stress As A Mediator Of Apoptosis. Immunology Today. 15(1): 7-10.
Wyllie, A. 1984. The Biology Of Cell Death In Tumours. Anticancer Research. 5(1): 131-136.
Cukierman, E., Pankov, R. and Yamada, K. M. 2002. Cell Interactions With Three-Dimensional Matrices. Current Opinion In Cell Biology. 14(5): 633-640.
Lee, J. M., Mhawech-Fauceglia, P., Lee, N., Parsanian, L. C., Lin, Y. G., Gayther, S. A. and Lawrenson, K. 2013. A Three-Dimensional Microenvironment Alters Protein Expression And Chemosensitivity Of Epithelial Ovarian Cancer Cells In Vitro. Laboratory Investigation. 93(5): 528-542.
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