ANTIOXIDANT AND CYTOTOXIC ACTIVITIES OF ARECA CATECHU SEED EXTRACT IN SWISS ALBINO MICE USING EAC CELL LINE IN DIFFERENT CULTURE MEDIUM
Objective: Areca nut is the dried ripe seeds of Areca catechu, belonging to family Palmae. Areca nut contains a number of alkaloids, belonging to pyridine and piperidine groups, derived from amino acid lysine. Arecoline a nicotinic acid-based alkaloid present which exerts sialagogue property. But the habit of chewing marketed gutka may cause oral leukoplakia, sometimes lead to squamous cell carcinoma. However, Based on the phytochemical compounds present, it is predicted that it must show antioxidant and may show cytotoxic activities.
Methods: The antioxidant activity (nitrite scavenging and hydrogen peroxide scavenging) and cytotoxic activity in two different medium were checked in EAC cell line using the swiss albino mice model.
Results: The study was also postulated an idea about the qualitative and quantitive analysis of Areca catechu. The plant extract showed good Nitrite and Hydrogen peroxide scavenging activity. The cytotoxicity study conducted in swiss albino mice, cell viability and IC50 value was checked. The cytotoxic activity of different concentrations of different fractions of plant extract was checked in two different medium, i.e., PBS and RPMI 1640. IC50 values for following fractions which were studied in PBS; as, 91.73±73 (µg/ml), 183±36.24 (µg/ml), 53.74±1.562 (µg/ml) for crude ethanolic extract, alkaloid fraction and flavonoid fraction respectively. IC50 value in RPMI 1640 medium obtained as; 44.18±1.09µg/ml, 54.27±0.2279µg/ml and 51.24±2.461µg/ml for crude ethanolic extract, Alkaloid and Flavonoid fraction, respectively.
Conclusion: Areca nut extract showed good scavenging activity depending on concentration. Relatively RPMI 1640 medium showed better cytotoxic activity than other mediums.
2. Boucher BJ, Mannan N. Metabolic effects of the consumption of Areca catechu. Addict Biol 2002;7:103–10.
3. Giri S, Idle JR, Chen C, Zabriskie TM, Krausz KW, Gonzalez FJ. A metabolonomic approach to the metabolism of the areca nut alkaloids arecoline and arecaidine in the mouse. Chem Res Toxicol 2006;19:818–27.
4. Chu NS. Neurological aspects of areca and betel chewing. Addict Biol 2002;7:111–4.
5. Shubhangi Mhaske, T Raju Ragavendra, CBS Dangi. Areca nut as a chemical carcinogen in oral squamous cell carcinoma-a review. Biomed Pharmacol J 2009;2:437-40.
6. Preetee Jaiswal, Pradeep kumar, VK Singh, DK Singh. Areca catechu L.: a valuable herbal medicine against different health problems. Res J Med Plants 2011;5. Doi:10.3923/rjmp.2011.145.152
7. Peter M, Som MD. Lymph Nodes of the Neck. Radiology 1987;165:593-600.
8. Barnes L, Organization WHO, Cancer IAfRo. Pathology and genetics of head and neck tumours. World Health Organization; 2005.
9. Mehmet O, Isik DK, Ibrahim HK, Muhammed EG. Ehrlich ascites carcinoma. Afr J Biotechnol 2011;10:2375-8.
10. Vogel AR, Tatchell BS, Furnis A, J Hannaford. Vogel's textbook of practical organic chemistry. 5th Edition by A. I. P. W. G. Smith; 1989.
11. Fair JD, Kormos CM. Tables showing the thickness value of commercial regular and preparative TLC plates. J Chromatogram 2008;1211:49-54.
12. Jonathan M Stoddard, Lien Nguyen, Hector Mata Chavez, Kelly Nguyen. TLC plates as a convenient platform for solvent-free reactions. Chem Commun 2007;12:1240–41.
13. Aromdee C, Panuwougse S, Anorach R, Vorarat S. A high pressure liquid chromatographic method for the determination of arecoline in Areca nut, Thai. J Pharm Sci 2003;27:41-7.
14. Azeez S, Amudhan S, Adiga S, Rao N, Laxminarayana A. Wound healing profile of Areca catechu extracts on different wound models in wistar rats. Kuwait Med J 2007;39:48-52.
15. Dar A, Khatoon S. Behavioural and biochemical studies of dichloromethane fraction from the Areca catechu nut. Pharmacol Biochem Behave 2000;65:1-6.
16. Dhinegra D, Sharma A. A review on antidepressant plants, nature products radiance. New Delhi: National Institute of Science Communication Information Resources 2006;5:146-68.
17. Dua J, Prasad DN, Tripathi AC, Gupta R. Role of traditional medicine in Neuro psychopharmacology, Asian J Pharm Clin Res 2009;2:42-6.
18. Chu NS. Sympathetic response to betel chewing. J Psychoact Drugs 1995;5:183-6.
19. Ingole SR, Rajput SK, Sharma SD. Cognition enhancers: current strategies and future perspectives. CRIPS 2008;9:42-6.
20. Lalithkumari H, Sirsi M. Antibacterial antifungal activity of Areca catechu Linn. Indian J Exp Biol 1965;7:66.
21. Nadkarni AK. Indian Material Medica. 3rd Ed, Mumbai; Bombay Popular Prakashan. Vol. 1. Parle M, Vasudevan M. Memory Enhancing Activity of Abana: An Indian Ayurvedic Poly-herbal Formulation. J Health Sci 2007;53:43-52.
22. Pithuyanukul P, Nithitanakal S, Bavovada R. Hepatoprotective potential of extracts from seeds of Areca catechu and nutgalls of quercus infectoria. Molecules 2009;14:4987-5000.
23. Reena R, Authikat N, Michael A. Study on the Areca nut for its antimicrobial properties. Pharmacognosy 2009;1:42-5.
24. Dashora N, Sodde V, Bhagat J, Prabhu KS, Lobo R. Antitumor activity of dendrophthoe falcate against ehrlich ascites carcinoma in swiss albino mice. Pharm Crops 2010;2:1-7.
25. Slinkard J, Singleton VL. Total phenol analysis: automation and comparison with manual methods. Am J Enol Vitic 1977;28:49-55.
26. Harinder PS. Quantification of tannins in tree and shrub foliage. A Laboratory Manual, Spinger-Science+Business media; 2003.
27. Vijay DT, Rajendra SB. Estimation of total phenol, tannin, alkaloid and flavonoid in hibiscus tiliaceus linn. wood extracts. Res Rev: J Pharmacogn Phytochem 2014;2:41-7.
28. Zhishen J, Mengcheng T, Jianming W. The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chem 1999;64:555-9.
29. Francis MA, Andrew WV. Antioxidant activity, nitric oxide scavenging activity and phenolic contents of Ocimum gratissimum leaf extract. J Med Plants Res 2010;4:2479-87.
30. Marcocci L, Maguire JJ, Droy Lefaix MT, Packer L. The nitric oxide scavenging properties of Ginkgo biloba extract. Biochem Biophys Res Commun 1994;201:748-55.
31. Gries Stefan. Some current quantitative problems in corpus linguistics and a sketch of some solutions. Language Linguistics 2015;16:93-117.
32. Babu BH, Shylesh BS, Padikkala J. Antioxidant and hepatoprotective effect of acanthus ilicifolius. Fitoterapia 2001;72:272-7.
33. Aline AB, Michel MM, Margareth LA. Technical evaluation of antioxidant activity. Med Chem 2014;4:7.
34. Zhang XY. Principles of chemical analysis. Beijing: China Science Press; 2000. p. 275.
35. Sneha M, Ankit KT, Mohsen S, Perumal AD, Shruti MK, Anwit SP, et al. Subcutaneous ehrlich ascites carcinoma mice model for studying cancer-induced cardiomyopathy. Sci Rep 2018;8:5599.
36. Mariana FM, Polyana LB, Aline CB, Frederico SM, Jose RP, Marize CV. Investigation of Ehrlich ascites tumor cell death mechanisms induced by Synadenium umbellatum Pax. J Ethnopharmacol 2012;139:319-29.
37. Terry LR, Richard AM, Andrew LN, Sarah D, Helene AB, Tracy JW, et al. Cell viability assays. Assay Guidance Manual; 2013. p. 2-11.
38. Soundararajan V, Sreenivasan S. Cytotoxicity of methanol extracts of Elaeis guineensis on MCF-7 and vero cell lines. Asian Pac J Trop Biomed 2012;2:826-9.
39. Nowroji K, Rahmah N, Kit C, Sreenivasan S. Cytotoxicity activity of root extract/fractions of Eurycoma longifolia Jack root against vero and Hs27cells. J Med Plants Res 2010;4:2383-7.
40. Robak J, Gryglewski RJ. Flavonoids are scavengers of superoxide anions. Biochem Pharmacol 1988;37:837-41.
41. Brown F, Mire SA. Developments in biologicals. The design and analysis of potency assays for biotechnology products. Geneve: Karger AG Medecine et Hygiene; 2002. p. 107.
42. Hayon T, Dvilansky A, Sphilberg O, Nathan I. Appraisal of the MTT based assay as a useful tool for predicting drug chemosensitivity in leukemia. Leukemia Lymphoma 2003;44:1957-62.
43. Bagan J, Sarrion G, Jimenez Y. Oral cancer: clinical features. Oral Oncol 2010;46:414-7.
44. Yellowitz JA, Horowitz AM, Drury TF, Goodman HS. Survey of U. S. dentists’ knowledge and opinions about oral pharyngeal cancer. J Am Dent Assoc 2000;131:653-61.
45. Rivera C, Venegas B. Histological and molecular aspects of oral squamous cell carcinoma (review). Oncol Lett 2014;8:7-11.
46. Gonzalez MA, Ruiz AI, Gil MA, Esteban F, Bravo M. Analysis of Ki-67 expression in oral squamous cell carcinoma: why Ki-67 is not a prognostic indicator. Oral Oncol 2010;46:525-30.
47. Verma R, Singh A, Jaiswal R, Chandra A, Verma R, Tak J. Association of Ki-67 antigen and p53 protein at invasive tumor front of oral squamous cell carcinoma. Indian J Pathol Microbiol 2014;57:553-7.
This work is licensed under a Creative Commons Attribution 4.0 International License.