IN VITRO ANTIOXIDANT POTENTIAL, FREE RADICAL SCAVENGING AND CYTOTOXIC ACTIVITY OF SIMAROUBA GLUACA LEAVES.
Keywords:Simarouba gluaca, Phenolics, Tannins, DPPH, FRAP, Reducing power, Iron chelation, Cytotoxicity
Objective: To investigate the total phenolic, flavonoids, tannin contents of Simarouba glauca leaves and evaluate the antioxidant potential and cytotoxic activity in different human cell lines.
Methods: The methanol, ethanol and water extracts of S. glauca leaves were examined for total phenolics, flavonoid and tannin content. Its antioxidant properties using FRAP, Phosphomolybdenum, Ferric ferrozine assays and free radical scavenging using DPPH were determined. Further, the reducing power and iron chelating effect of the extract using spectrophotometric assays were estimated. Finally, the cytotoxic activity against few human cancer cell lines were also examined using MTT assay.
Results: The phytochemical investigations revealed that S. glauca leaves has only 0.14 to 0.18% of flavonoids, 250-400Âµg/mg phenolics and 67-200Âµg/mg tannin content in various solvent extracts. The extracts exhibited good reducing power with similar EC50 values aproximately in the range of 57-61Âµg /ml. Further, the leaf extracts showed iron chelation effect which was more pronounced in aqueous extract with IC50 value of 332Âµg/ml and exhibited very strong DPPH radical scavenging activity with IC50 vlaues ranging from 9-13Âµg/ml in various extracts. The methanolic extracts showed good antioxidant potential using FRAP and phopshomolybdenum methods, whereas the aqueous extract exihibited more pronounced antioxidant activty using sensitive ferric ferrozine assay with 580Âµg AAE/mg extract. The methanolic extract showed strong cytotoxic effect on SCC9 cancer cell line and less potent on HCT116 cancer cells.
Conclusion: The findings suggests that Simarouba glauca leaf extract contain bioactive molecules which exhibit antioxidant activity that could be synergisticaly influencing the cytotoxic activity in selected cancer cell lines.
Halliwell B, Gutteridge J. Free Radicals in Biology and Medicine, 4 Ed. Oxford University Press: Oxford; 2007.
Rice-Evans CA, Miller N, Paganga G. Antioxidant properties of phenolic compounds. Trends Plant Sci 1997;2:152-9.
Neergheen VS, Bahorun T, Taylor EW, Jen L, Aruoma O. Targeting specific cell signaling transduction pathways by dietary and medicinal phytochemicals in cancer chemoprevention. Toxicol 2010;278(2):229â€“41.
Fresco P, Borges F, Marques MPM, Diniz C. The anticancer properties of dietary polyphenols and its relation with apoptosis. Cur Pharm Design 2010;16(1):114â€“34.
Huang WY, Cai YZ, Zhang YB. Natural phenolic compounds from medicinal herbs and dietary plants: potential use for cancer prevention. Nut Cancer An Int J 2010;62(1):1â€“20.
Moron J, Merrien MA, Polonsky J. Sur la biosynthe`se des quassinoÄ±Â¨des de Simaruba glauca (Simarubaceae). Phytochem 1971;10:585.
Patel DP, Verma VD, Loknathan TR, Bhatt KC, Mishra JB. Simarouba glaucaâ€“a non traditional oil source. Ind Farm 1997;47:7-9.
Franssen FFJ, Smeijsters LJJW, Berger I, Aldana BEM. In vivo and in vitro antiplasmodial activities of some plants traditionally used in guatemala against malaria. Antimicr Agents Chem 1997;41(7):1500â€“3.
Valdes AF, MartÃnez JM, Lizama RS, Paul Cos MV, Maes L. In vitro antimicrobial activity of the Cuban medicinal plants Simarouba glauca, Melaleuca leucadendron and Artemisia absinthium. Mem Inst Oswaldo Cruz Rio de Janeiro 2008;103(6):615-8.
Mikawlrawng K, Sandeep Kaushik, Pushker AK, Suresh Kumar, Kameshwor Singh M, Sharma GS. Comparative in vitro antifungal activities of Simarouba glauca against Fusarium oxysporum and Aspergillus parasiticus. J Med Plant Stud 2014;2(3):1-7.
Lidia MG, Fereire V, Alonzo A, Caceres A. Ethnobotanical survey of the medicinal flora used by the caribs of guatemala. J Ethnopharmacol 1991;34:173â€“87.
Monjour L, Rouquier F, Alfred C, Polonsky J. Therapeutic trials of experimental murine malaria with the quassinoid, glaucarubinone Comptes Rendus de l'Academie des. Sci-Serie III 1987;304(6):129-32.
Yeo D, Nhi Huynh, Beutler JA, Christophi C, Shulkes A, Baldwin GS, et al. Glaucarubinone and gemcitabine synergistically reduce pancreatic cancer growth via down-regulation of P21-activated kinases. Cancer Let 2014:346:264â€“72.
Zarse K, Bossecker A, MÃ¼ller-Kuhrt L, Siems K, Hernandez MA, Berendsohn WG, et al. The phytochemical glaucarubinone promotes mitochondrial metabolism, reduces body fat, and extends lifespan of Caenorhabditis elegans. Horm Metab Res 2011;43:241â€“3.
Rivero-Cruz JF, Lezutekong R, Lobo-Echeverri T, Ito A, Mi Q, Chai H, et al. Cytotoxic constituents of the twigs of Simarouba glauca collected from a plot in Southern Florida. Phyt Res 2005;19(2):136-40.
Zhao H, Dong J, Lu J, Chen J, Li Y, Shan L, et al. Effects of extraction solvent mixtures on antioxidant activity evaluation and their extraction capacity and selectivity for free phenolic compounds in barley (Hordeum vulgare L.). J Agric Food Chem 2006;54(19):7277-86.
Boeing JS, BarizÃ£o EO, Costa e Silva B, Montanher PF, Almeida VC, Visentainer JV. Evaluation of solvent effect on the extraction of phenolic compounds and antioxidant capacities from the berries: application of principal component analysis. Chem Centr J 2014;8:48.
Akowuah GA, Ismail Z, Norhayati I, Sadikun A. The effects of different extraction solvents of varying polarities on polyphenols of Orthosiphon stamineus and evaluation of the free radical-scavenging activity. Food Chem 2005;93(2):311-7.
Umesh TG, Abhirami D. Evaluation of antioxidant potential and reducing power of callus induced from leaves of Asystasia gangetica (L.) T Anderson Int J Pharm Pharm Sci 2014;6(8):532-8.
Sun B, Richardo-Da-Silvia JM, Spranger I. Critical factors of vanillin assay for catechins and proanthocyanidins. J Agri Food Chem 1998;46:4267-74.
Berker KI, Guclu K, Demirata B, Apak R. A novel antioxidant assay of ferric reducing capacity measurement using ferrozine as the colour forming complexation reagent. Anal Methods 2010;2:1770â€“8.
Umesh TG. In vitro callus induction and antioxidant potential of Decalepis hamiltonii (wight and arn). Int J Pharm Pharm Sci 2014;6(6):452-6.
Benzie IFF, Strain JJ. The ferric reducing ability of plasma (FRAP) as a measure of antioxidant power: the FRAP assay. Anal Biochem 1996;239:70-6.
Prieto P, Pineda M, Aguilar M. Spectrophotometric Quantitation of antioxidant capacity through the formation of a phosphomolybdenum complex: specific application to the determination of vitamin E. Anal Biochem 1999;269:337-41.
Dinis TCP, Madeira VMC, Almeida LM. Action of phenolic derivatives (acetaminophen, salicylate and 5-aminosalicylate) as inhibitors of membrane lipid peroxidation and as peroxyl radical scavengers. Arch Biochem Biophys 1994;315:161-9.
Lakshmi KS, Sangeetha D, Sivamani S, Tamilarasan M, Rajesh TP, Anandraj B. In vitro antibacterial, antioxidant, haemolytic, thrombolytic activities and phytochemical analysis of Simarouba glauca leaves extracts. Int J Pharm Sci Res 2014;5(2):432-7.
Bravo L. Polyphenols: chemistry, dietary sources, metabolism and nutritional significance. Nutr Rev 1998;56:317-33.
Fresco P, Borges F, Diniz C, Marques MPM. New insights on the anticancer properties of dietary polyphenols. Med Res Rev 2006;26(6):747â€“66.
Van Acker SABE, Berg DVD, Tromp MNJL, Griffioen DH, Van Bennekom WP, Van Der Vijgh WJF, et al. Structural aspects of antioxidant activity of flavonoids. Free Rad Bio Med 1996;20(3):331â€“42.
Ruch RJ, Cheng SJ, Klaunig JE. Prevention of cytotoxicity and inhibition of Intercellular communication by antioxidant catechins isolated from Chinese green tea. Carcinogens 1989;10:1003-8.
Hagerman AE, Riedl KM, Alexander Jones G, Sovik KN, Ritchard NT, Hartzfeld PW, et al. High molecular weight plant polyphenolics (Tannins) as biological antioxidants. J Agric Food Chem 1998;46:1887âˆ’92.
Koleckar V, Kubikova K, Rehakova Z, Kuca K, Jun D, Jahodar L, et al. Condensed and hydrolysable tannins as antioxidants influencing the health. Mini Rev Med Chem 2008;8:436-47.
Gulcin I, Huyut Z, Elmastas M, HassanY, Aboul-Enein. Radical scavenging and antioxidant activity of tannic acid. Arab J Chem 2010;3(1):43â€“53.
Chung K, Yee Wong T, Wei C, Huang Y, Lin Y. Tannins and Human Health: a review. Cri Rev Food Sci Nut 1998;38(6):421â€“64.
Maranhao HML, Vasconcelos CFB, Rolim LA, Rolim Neto PJ, Silva Neto J, da Silva Filho R, et al. Hepatoprotective effect of the aqueous extract of Simarouba amara Aublet (Simaroubaceae) stem bark against carbon tetrachloride (CCl4)-induced hepatic damage in rats. Mol 2014;19:17735-46.
Scalbert A, Manach C, Morand C, Remsy C. Dietary of polyphenols and the prevention of diseases. Cri Rev Food Sci Nut 2005;45:287â€“306.
Rahman A, Kim E, Kang S. Antibacterial and antioxidant properties of Ailanthus altissima swingle leave extract to reduce foodborne pathogens and spoiling bacteria. J Food Saf 2009;29:499â€“510.
Albouchi F, Hassen I, Casabianca H, Hosni K. Phytochemicals, antioxidant, antimicrobial and phytotoxic activities of Ailanthus altissima (Mill.) Swingle leaves. S Afr J Bot 2013;87:164â€“74.
Nonita PP, Mylene MU. Antioxidant and cytotoxic activities and phytochemical screening of four Philippine medicinal plants. J Med Plants Res 2010;4(5):407-14.
Siddhuraju P, Becker K. The antioxidant and free radical scavenging activities of processed cowpea (Vigna unguiculata (L.) Walp.) seed extracts. Food Chem 2007;101:10â€“9.
Amarowicz R, Pegg RB, Rahimi-Moghaddam P, Barl B, Weil JA. Free-radical scavenging capacity and antioxidant activity of selected plant species from the Canadian prairies. Food Chem 2004;84:551â€“62.
Sugihara N, Ohnishi M, Imamura M, Furuno K. Differences in antioxidative efficiency of catechins in various metal-induced lipid peroxidations in cultured hepatocytes. J Health Sci 2001;47:99â€“106.
Brown JE, Khodr H, Hider RC, Rice-Evans CA. Structural dependence of flavonoid interactions with Cu2+ ions: implications for their antioxidant properties. Biochem J 1998;330:1173â€“8.
Kaliora AC, Kogiannou DAA, Kefalas P, Papassideri IS, Kalogeropoulos N. Phenolic profiles and antioxidant and anticarcinogenic activities of Greek herbal infusions; balancing delight and chemoprevention? Food Chem 2014;142:233â€“41.
Chinery R, Brockman JA, Peeler MO, Yu Shyr, Beauchamp RD, Robert JC. Antioxidants enhance the cytotoxicity of chemotherapeutic agents in colorectal cancer: a p53-independent induction of p21WAF1/CIP1 via C/EBP . Nat Med 1997;3:1233â€“41.
Houel E, Stien D, Bourdy G, Deharo E. Quassinoids: anticancer and antimalarial activities. In: KG Ramawat, JM MeÂ´rillon. Editors. Natural Products. Berlin Heidelberg Springer-Verlag; 2013. p. 3775-802.
Mesquita ML, Paula J, Pessoa C, de Moraes M, Costa-Lotufo LV, Grougnet R, et al. Cytotoxic activity of brazilian cerrado plants used in traditional medicine against cancer cell lines. J Ethnopharmacol 2009;123:439â€“45.
Suffness M, Pezzuto JM. Assays related to cancer drug discovery. In: Hostettmann K, Edited. Methods in Plant Biochemistry: Assays for Bioactivity. London: 6. Academic Press; 1990. p. 71-133.