LEAVES OF ANDROGRAPHIS PANICULATA IS AN ANTIOXIDANT AND ANTICANCER AGENT
Objective: Andrographis paniculata (Family: Acanthaceae) is a well-known medicinal plant used in the Indian traditional system of medicine for the treatment of many chronic diseases. The present study was aimed to quantify secondary metabolites, determine antioxidant, and anticancer activity of ethanol extract of A. paniculata leaves.
Methods: Leaf sample was macerated with ethanol solvent. Alkaloids, terpenoids, saponins, phenols, and flavonoids were quantified with standard calibrations. The antioxidant potential was tested using 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), and ferric reducing antioxidant power (FRAP) assays. In vitro anticancer activity was evaluated using human epithelial type 2 (HEp-2) cell line. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to estimate the cytotoxicity of the extracts. Apoptotic and necrotic effects were characterized by DNA fragmentation assay and fluorescence microscopy using the dual acridine orange/ethidium bromide (AO/EB) staining method.
Results: The phytochemical analysis reveals the presence of alkaloids, saponins, phenols, flavonoids, terpenoids, and steroids. Alkaloids, terpenoids, saponins, phenol, and flavonoid content were recorded as follows: 9.84%, 8.42%, 13.94%, 44.37 mg gallic acid equivalent/100 g, and 904 mg quercetin equivalent/100 g, respectively. The antioxidant activity from DPPH, ABTS, and FRAP assays showed dose-dependent inhibition of free radicals. In cell viability tests, cell death with increasing extract concentration was observed. DNA fragmentation and AO/EB stain confirmed apoptosis and necrosis in extract-treated cells.
Conclusion: The results indicate that A. paniculata is a promising source for the development of antioxidant and anticancer drugs.
2. Pham-Huy LA, He H, Pham-Huy C. Free radicals, antioxidants in disease and health. Int J Biomed Sci 2008;4:89-96.
3. Lobo V, Patil A, Phatak A, Chandra N. Free radicals, antioxidants and functional foods: Impact on human health. Pharmacogn Rev 2010;4:118-26.
4. Tagne RS, Telefo BP, Nyemb JN, Yemele DM, Njina SN, Goka SM, et al. Anticancer and antioxidant activities of methanol extracts and fractions of some Cameroonian medicinal plants. Asian Pac J Trop Med 2014;7S1:S442-7.
5. Kumar A, Selvakumar S. Antiproliferative efficacy of Tabernaemontana divaricata against HEp-2 cell line and Vero cell line. Pharmacogn Mag 2015;11 Suppl 1:S46-52.
6. Desai AG, Qazi GN, Ganju RK, El-Tamer M, Singh J, Saxena AK, et al. Medicinal plants and cancer chemoprevention. Curr Drug Metab 2008;9:581-91.
7. Meybodi NM, Mortazavian AM, Monfared AB, Sohrabvandi S, Meybodi FA. Phytochemicals in cancer prevention: A review of the evidence. Int J Cancer Manag 2017;10:e7219.
8. Akbar S. Andrographis paniculata: A review of pharmacological activities and clinical effects. Altern Med Rev 2011;16:66-77.
9. Rajalakshmi V, Cathrine L. Phytochemical screening and antimicrobial activity of ethanolic extract of Andrographis paniculata. J Pharmacogn Phytochem 2016;5:175-7.
10. Patel PR, Nagar AA, Patel RC, Rathod DK, Patel VR. In vitro anticancer activity of Rubia cordifolia against HeLa and HEp-2 cell lines. Int J Pharm Pharm Sci 2011;3 Suppl 2:70-1.
11. Ramesh S, Dilipan E, Mayavu P. Effects of drugs against antioxidant and cytotoxic (HEp-2 cell line) activity compound from marine animals Conus amadis venom (Gmelin, J.F, 1791). Int J Pharm Pharm Sci 2014;6:638-43.
12. Handa SS. An overview of extraction techniques for medicinal and aromatic plants. In: Handa SS, Khanuja SP, Longo G, Rakesh DD, editors. Extraction Technologies for Medicinal and Aromatic Plants. Trieste: ICS-UNIDO; 2008. p. 21-54.
13. Trease GE, Evans WC. Pharmacognosy. 12th ed. London: Balliere Tindall; 1983.
14. Demiray S, Pintado ME, Castro PM. Evaluation of phenolic profiles and antioxidant activities of Turkish medicinal plants: Tilia argentea, Crataegi folium leaves and Polygonum bistorta roots. World Acad Sci Eng Technol 2009;54:312-7.
15. Chang CC, Yang MH, Wen HM, Chern JC. Estimation of total flavonoid content in Propolis by two complementary colorimetric methods. J Food Drug Anal 2002;10:178-82.
16. Obadoni BO, Ochuko PO. Phytochemical studies and comparative efficacy of the crude extracts of some haemostatic plants in Edo and Delta states of Nigeria. Glob J Pure Appl Sci 2002;8:203-8.
17. Harborne JB. Phytochemical Methods: A Guide to Modern Techniques of Plant Analysis. London: Chapman and Hall; 1973.
18. Ferguson NM. A Textbook of Pharmacognosy. New York: The Macmillon Company; 1956. p. 191.
19. Braca A, Sortino C, Politi M, Morelli I, Mendez J. Antioxidant activity of flavonoids from Licania licaniaeflora. J Ethnopharmacol 2002;79:379-81.
20. Arnao M, Cano A, Acosta M. The hydrophilic and lipophilic contribution to total antioxidant activity. Food Chem 2001;73:239-44.
21. Oyaizu M. Studies on products of browning reaction: Antioxidative activity of products of browning reaction prepared from glucosamine. Jpn J Nutr Diet 1986;44:307-15.
22. Mosmann T. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immunol Methods 1983;65:55-63.
23. Liu K, Liu PC, Liu R, Wu X. Dual AO/EB staining to detect apoptosis in osteosarcoma cells compared with flow cytometry. Med Sci Monit Basic Res 2015;21:15-20.
24. El-Jemli M, Kamal R, Marmouzi I, Zerrouki A, Cherrah Y, Alaoui K. Radical-scavenging activity and ferric reducing ability of Juniperus thurifera (L.), J. oxycedrus (L.), J. phoenicea (L.) and Tetraclinis articulata (L.). Adv Pharmacol Sci 2016;2016:6392656.
25. Das P, Srivastav AK. Phytochemical extraction and characterization of the leaves of Andrographis paniculata for its anti-bacterial, anti-oxidant, anti-pyretic and anti-diabetic activity. Int J Innov Res Sci Eng Technol 2014;3:15176-84.
26. Nagajothi S, Mekala P, Raja A, Raja MJ, Senthilkumar P. Andrographis paniculata: Qualitative and quantitative phytochemical analysis. J Pharmacogn Phytochem 2018;7:1251-3.
27. Chintalapani S, Swathi MS, Narasu ML. Phytochemical screening and in vitro antioxidant activity of whole plant extracts of Sesuvium portulacastrum L. Asian J Pharm Clin Res 2018;11:322-7.
28. Jassal PS, Sharma M. Evaluation of antioxidant, antibacterial, antihemolytic, and phytochemical properties of Ficus benjamina, Ficus infectoria and Ficus krishnae. Asian J Pharm Clin Res 2019;12:68-73.
29. Liang N, Kitts D. Antioxidant property of coffee components: Assessment of methods that define mechanisms of action. Molecules 2014;19:19180-208.
30. Sangeetha S, Archit R, Mythili S, Sathiavelu A. A detailed analysis of the antioxidant activity of the medicinal plant Andrographis paniculata. Int J Drug Dev Res 2014;6:231-8.
31. Ratnavathi CV, Komala VV. Sorghum grain quality. In: Ratnavathi CV, Patil JV, Chavan UD, editors. Sorghum Biochemistry: An Industrial Perspective. London: Elsevier; 2016. p. 1-61.
32. Adeoye BO, Akinrinde AS, Oyagbemi AA, Omobowale TO, Afolayan AJ, Adedapo AA. Phytochemical profiling, antioxidant activities and essential oil constituents of Andrographis paniculata. Trop Vet 2018;36:52-64.
33. Cerretani L, Bendini A. Rapid assays to evaluate the antioxidant capacity of phenols in virgin olive oil. In: Preedy VR, Watson RR, editors. Olives and Olive Oil in Health and Disease Prevention. London: Elsevier; 2010. p. 625-35.
34. Prakash SE, Ali SH, Divya N, Rani RV, Manavalan R. Evaluation of in vitro antioxidant activity of leaf extract of Andrographis paniculata. Res J Pharm Biol Chem Sci 2011;2:891-5.
35. Adedapo AA, Adeoye BO, Sofidiya MO, Oyagbemi AA. Antioxidant, antinociceptive and anti-inflammatory properties of the aqueous and ethanolic leaf extracts of Andrographis paniculata in some laboratory animals. J Basic Clin Physiol Pharmacol 2015;26:327-34.
36. Riss TL, Moravec RA, Niles AL, Duellman S, Benink HA, Worzella TJ, et al. Cell viability assays. In: Sittampalam GS, Grossman A, Brimacombe K, Arkin M, Auld D, Austin CP, et al, editors. Assay Guidance Manual. Bethesda, MD: Eli Lilly and Company and the National Center for Advancing Translational Sciences; 2004.
37. Amudha CK, Deeba F, Rajarajan P. Characterization and anti-neoplastic potential of phytofabricated silver nanoparticles on human breast cancer cell line (MCF-7). Int J Appl Pharm 2019;11:70-7.
38. Danial NN, Korsmeyer SJ. Cell death: Critical control points. Cell 2004;116:205-19.
39. Gozuacik D, Kimchi A. Autophagy as a cell death and tumor suppressor mechanism. Oncogene 2004;23:2891-906.
40. Proskuryakov SY, Konoplyannikov AG, Gabai VL. Necrosis: A specific form of programmed cell death? Exp Cell Res 2003;283:1-16.
41. Saadat YR, Saeidi N, Vahed SZ, Barzegari A, Barar J. An update to DNA ladder assay for apoptosis detection. Bioimpacts 2015;5:25-8.
42. Goodman L. The kiss of death. J Clin Invest 2004;113:1662.
43. Mao W, He P, Wang W, Wu X, Wei C. Andrographolide sensitizes HEp-2 human laryngeal cancer cells to carboplatin-induced apoptosis by increasing reactive oxygen species levels. Anticancer Drug 2019;30:e0774.
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