Antioxidant and cytotoxic activity of stems of Smilax zeylanica in vitro

Mohammad Nasir Uddin; Taksim Ahmed; Sanzida Pathan; Md. Mamun Al-Amin; Md. Sohel Rana

doi:10.1515/jbcpp-2014-0114

Article

Antioxidant and cytotoxic activity of stems of Smilax zeylanica in vitro

Mohammad Nasir Uddin , Taksim Ahmed , Sanzida Pathan , Md. Mamun Al-Amin and Md. Sohel Rana

Published/Copyright: April 22, 2015

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From the journal Journal of Basic and Clinical Physiology and Pharmacology Volume 26 Issue 5

Abstract

Background: Plant-derived phytochemicals consisting of phenols and flavonoids possess antioxidant properties, eventually rendering a lucrative tool to scavenge reactive oxygen species. This study was carried out to evaluate in vitro antioxidant and cytotoxic potential of methanolic extract and petroleum ether extracts of Smilax zeylanica L. stems.

Methods: Phytochemical screening was done following standard procedures. Antioxidant activity was tested using several in vitro assays, viz., 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay, NO assay, H₂O₂ assay, CUPRAC assay, FRAP assay and total antioxidant capacity assay. Total phenol and flavonoid contents were determined by colorimetric method. Brine shrimp lethality and MTT cell viability assays were used for cytotoxic potential.

Results: Preliminary phytochemical study revealed the presence of flavonoids and glycosides in both extracts. Methanolic extract was found to possess stronger antioxidant potential than petroleum ether extracts in all assays. The IC₅₀ value of methanolic extract was 29.14±0.39 μg/mL, 120.30±3.32 μg/mL and 78.41±5.53 μg/mL in DPPH assay, NO assay and H₂O₂ assay, respectively. Likewise, total phenol [56.78 mg/g gallic acid (GAE)] and flovonoid [125.69 mg/g quercetin equivalents (QE)] were higher in methanolic extract. In cytotoxicity assays, petroleum ether extract showed stronger activity in both brine shrimp lethality (LC₅₀ 2.85±0.13 μg/mL) and MTT cell viability assay (IC₅₀ 15.49±1.18 μg/mL).

Conclusions: These findings demonstrate that methanolic extracts could be considered as potential sources of natural antioxidant, whereas petroleum ether extracts could be explored for promising anticancer molecules.

Keywords: antioxidant activity; brine shrimp; DPPH; H2O2; MCF7; MTT; Smilax zeylanica

Corresponding author: Mohammad Nasir Uddin, Department of Pharmaceutical Sciences, North South University, Dhaka, Bangladesh, Phone: +8801715824327, E-mail: nasiruddin@northsouth.edu; nasiruddin1286@gmail.com. http://orcid.org/0000-0002-5668-5374; and Laboratory of Natural Products Research, Department of Pharmacy, Jahangirnagar University, Dhaka, Bangladesh

Acknowledgments

The authors would like to thank native English speaker, Mr. Jason Chau, School of Pharmacy, University of Waterloo, Ontario, Canada, for critically revising the English of the manuscript, and Mohammad Ehsanul Haque Mazumder, Discipline of Biomedical Science, School of Medical Sciences, The University of Sydney, Australia, for his valuable information, suggestions and critical reading of the manuscript.

Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

Research funding: None declared.

Employment or leadership: None declared.

Honorarium: None declared.

Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

References

1. Sies H. Oxidative stress: oxidants and antioxidants. Exper Physiol 1997;82:291–5.10.1113/expphysiol.1997.sp004024Search in Google Scholar

2. Halliwell B, Gutteridge JM. Oxygen toxicity, oxygen radicals, transition metals and disease. Biochem J 1984;219:1–14.10.1042/bj2190001Search in Google Scholar

3. Dillard CJ, German JB. Phytochemicals: nutraceuticals and human health. J Sci Food Agric 2000;80:1744–56.10.1002/1097-0010(20000915)80:12<1744::AID-JSFA725>3.0.CO;2-WSearch in Google Scholar

4. Ramarathnam N, Osawa T, Ochi H, Kawakishi S. The contribution of plant food antioxidants to human health. Trends Food Sci Tech 1995;6:75–82.10.1016/S0924-2244(00)88967-0Search in Google Scholar

5. Miliauskas G, Venskutonis PR, van Beek TA. Screening of radical scavenging activity of some medicinal and aromatic plant extracts. Food Chem 2004;85:231–7.10.1016/j.foodchem.2003.05.007Search in Google Scholar

6. Madhavan V, Hemalatha TH, Murali A, Yoganarasimhan SN. Antiepileptic activity of alcohol and aqueous extracts of roots and rhizomes and Smilax zeylanica Linn. Pharmacol Online 2008;3:263–72.Search in Google Scholar

7. Evans WC. Trease and Evans pharmacognosy, 15th ed. Edinburgh, New York: WB Saunders, 2002:300, 480.Search in Google Scholar

8. Kar DK, Sen S. Smilax zeylanica Linn. – a new source of diosgenin. Curr Sci 1984;53:661.Search in Google Scholar

9. Rajesh V, Perumal P, Sundarrajan T. Antidiabetic activity of methanolic extract of Smilax zeylanica Linn. in streptozotocin induced diabetic rats. Internet J Endocrinol 2010;6.10.5580/1d39Search in Google Scholar

10. Rajesh V, Perumal P, Chinthakindhi V, Prabhakaran S, Hymavathi G, Guntupalli T. In vitro evaluation of Smilax zeylanica Linn. leaves for anthelmintic activity. Internet J Pharmacol 2010;9.10.5580/797Search in Google Scholar

11. Murali A, Ashok P, Madhavan V. In vitro antioxidant activity and HPTLC studies on the roots and rhizomes of Smilax zeylanica L.(Smilacaceae). Int J Pharm Pharm Sci 2011;3:192–5.Search in Google Scholar

12. Thirugnanasampandan R, Mutharaian V, Bai VN. In vitro propagation and free radical studies of Smilax zeylanica Vent. Afr J Biotechnol 2009;8:395–400.Search in Google Scholar

13. Murali A, Ashok P, Madhavan V. Screening of methanol extract of roots and rhizomes of Smilax zeylanica L for hepatoprotective effect against carbontetrachloride induced hepatic damage. J Exp Integr Med 2012;2:237–44.10.5455/jeim.050512.or.028Search in Google Scholar

14. Bari M, Islam W, Khan A. Pesticidal activity of Smilax zeylanica L. extracts on Cryptolestes pusillus (Schon.)(Coleoptera: Cucujidae). J Bangladesh Acad Sci 2010;34:205–8.10.3329/jbas.v34i2.6868Search in Google Scholar

15. Ahmed T, Uddin MN, Ahmed SF, Saha A, Farhana K, Rana MS. In vitro evaluation of antioxidant potential of Artocarpus chama Buch fruits. J App Pharm Sci 2012;2:75–80.10.7324/JAPS.2012.21015Search in Google Scholar

16. Hasan R, Hossain M, Akter R, Jamila M, Mazumder EH, Islam I, et al. Antioxidant, antidiarrhoeal and cytotoxic properties of Punica granatum Linn. Latin Am J Pharm 2009;28:783–8.Search in Google Scholar

17. Ghani A. Medicinal plants of Bangladesh, 2nd ed. Dhaka, Bangladesh: The Asiatic Society of Bangladesh, 2003.Search in Google Scholar

18. Sadhu SK, Okuyama E, Fujimoto H, Ishibashi M. Separation of Leucas aspera, a medicinal plant of Bangladesh, guided by prostaglandin inhibitory and antioxidant activities. Chem Pharm Bull 2003;51:595–8.10.1248/cpb.51.595Search in Google Scholar PubMed

19. Braca A, De Tommasi N, Di Bari L, Pizza C, Politi M, Morelli I. Antioxidant principles from Bauhinia tarapotensis. J Nat Prod 2001;64:892–5.10.1021/np0100845Search in Google Scholar PubMed

20. Govindarajan R, Rastogi S, Vijayakumar M, Shirwaikar A, Rawat AK, Mehrotra S, et al. Studies on the antioxidant activities of Desmodium gangeticum. Biol Pharm Bull 2003;26:1424–7.10.1248/bpb.26.1424Search in Google Scholar PubMed

21. Ruch RJ, Cheng S-J, Klaunig JE. Prevention of cytotoxicity and inhibition of intercellular communication by antioxidant catechins isolated from Chinese green tea. Carcinogenesis 1989;10:1003–8.10.1093/carcin/10.6.1003Search in Google Scholar PubMed

22. Apak R, Güçlü K, Özyürek M, Karademir SE. Novel total antioxidant capacity index for dietary polyphenols and vitamins C and E, using their cupric ion reducing capability in the presence of neocuproine: CUPRAC method. J Agric Food Chem 2004;52:7970–81.10.1021/jf048741xSearch in Google Scholar PubMed

23. Oyaizu M. Studies on products of browning reactions. Antioxidative activities of products of browning reaction prepared from glucosamine. Jpn J Nutr 1986;44:307–15.10.5264/eiyogakuzashi.44.307Search in Google Scholar

24. 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;269:337–41.10.1006/abio.1999.4019Search in Google Scholar PubMed

25. Yu L, Haley S, Perret J, Harris M, Wilson J, Qian M. Free radical scavenging properties of wheat extracts. J Agric Food Chem 2002;50:1619–24.10.1021/jf010964pSearch in Google Scholar PubMed

26. Chang C, Yang M, Wen H, Chern J. Estimation of total flavonoids content in propolis by two complementary colormetric methods. J Food Drug Anal 2002;10:178–82.10.38212/2224-6614.2748Search in Google Scholar

27. Meyer BN, Ferrigni NR, Putnam JE, Jacobsen LB, Nichols DE, McLaughlin JL. Brine shrimp: a convenient general bioassay for active plant constituents. Planta Med 1982;45:31–4.10.1055/s-2007-971236Search in Google Scholar PubMed

28. McLaughlin JL. Bench-top bioassays for the discovery of bioactive compounds in higher plants. Brenesia 1991;34:1–14.Search in Google Scholar

29. Gil MI, Ferreres F, Tomás-Barberán FA. Effect of postharvest storage and processing on the antioxidant constituents (flavonoids and vitamin c) of fresh-cut spinach. J Agric Food Chem 1999;47:2213–7.10.1021/jf981200lSearch in Google Scholar PubMed

30. Kähkönen MP, Hopia AI, Vuorela HJ, Rauha J-P, Pihlaja K, Kujala TS, et al. Antioxidant activity of plant extracts containing phenolic compounds. J Agric Food Chem 1999;47:3954–62.10.1021/jf990146lSearch in Google Scholar

31. Shimada K, Fujikawa K, Yahara K, Nakamura T. Antioxidative properties of xanthan on the autoxidation of soybean oil in cyclodextrin emulsion. J Agric Food Chem 1992;40:945–8.10.1021/jf00018a005Search in Google Scholar

32. Yokozawa T, Chen CP, Dong E, Tanaka T, Nonaka G-I, Nishioka I. Study on the inhibitory effect of tannins and flavonoids against the 1,1-diphenyl-2-picrylhydrazyl radical. Biochem Pharmacol 1998;56:213–22.10.1016/S0006-2952(98)00128-2Search in Google Scholar

33. Moncada S, Palmer RM, Higgs EA. Nitric oxide: physiology, pathophysiology, and pharmacology. Pharmacol Rev 1991;43:109–42.10.1016/S0031-6997(25)06663-3Search in Google Scholar

34. Halliwell B. Reactive oxygen species in living systems: source, biochemistry, and role in human disease. Am J Med 1991;91:S14–22.10.1016/0002-9343(91)90279-7Search in Google Scholar

35. Tsutomu N, Munetaka Y, Toshihiko O, Shunro K. Suppression of active oxygen-induced cytotoricity by flavonoids. Biochem Pharmacol 1993;45:265–7.10.1016/0006-2952(93)90402-ISearch in Google Scholar

36. Xu J, Li X, Zhang P, Li Z-L, Wang Y. Antiinflammatory constituents from the roots of Smilax bockii warb. Arch Pharm Res 2005;28:395–9.10.1007/BF02977667Search in Google Scholar

37. Gülçin Đ, Elmastaş M, Aboul-Enein HY. Determination of antioxidant and radical scavenging activity of Basil (Ocimum basilicum L. Family Lamiaceae) assayed by different methodologies. Phytother Res 2007;21:354–61.10.1002/ptr.2069Search in Google Scholar

38. Gülçin I. Measurement of antioxidant ability of melatonin and serotonin by the DMPD and CUPRAC methods as trolox equivalent. J Enzyme Inhib Med Chem 2008;23:871–6.10.1080/14756360701626223Search in Google Scholar

39. Osawa T. Novel natural antioxidants for utilization in food and biological systems. In: Uritani I, Garcia V, Mendoza E, editors. Postharvest biochemistry of plant food-materials in the tropics. Tokyo, Japan: Japan Scientific Societies Press, 1994:241–51.Search in Google Scholar

40. Li J, Bi X, Zheng G, Hitoshi Y, Ikeda T, Nohara T. Steroidal glycosides and aromatic compounds from Smilax riparia. Chem Pharm Bull 2006;54:1451–4.10.1248/cpb.54.1451Search in Google Scholar

41. Cakir A, Mavi A, Yıldırım A, Duru ME, Harmandar M, Kazaz C. Isolation and characterization of antioxidant phenolic compounds from the aerial parts of Hypericum hyssopifolium L. by activity-guided fractionation. J Ethnopharmacol 2003; 87:73–83.10.1016/S0378-8741(03)00112-0Search in Google Scholar

42. Ko FN, Cheng ZJ, Lin CN, Teng CM. Scavenger and antioxidant properties of prenylflavones isolated from Artocarpus heterophyllus. Free Rad Biol Med 1998;25:160–8.10.1016/S0891-5849(98)00031-8Search in Google Scholar

43. Moridani MY, Pourahmad J, Bui H, Siraki A, O’Brien PJ. Dietary flavonoid iron complexes as cytoprotective superoxide radical scavengers. Free Rad Biol Med 2003;34:243–53.10.1016/S0891-5849(02)01241-8Search in Google Scholar

44. Kuo Y-H, Hsu Y-W, Liaw C-C, Lee JK, Huang H-C, Kuo L-MY. Cytotoxic phenylpropanoid glycosides from the stems of Smilax china. J Nat Prod 2005;68:1475–8.10.1021/np050109qSearch in Google Scholar PubMed

45. Ivanova A, Mikhova B, Batsalova T, Dzhambazov B, Kostova I. New furostanol saponins from Smilax aspera L. and their in vitro cytotoxicity. Fitoterapia 2011;82:282–7.10.1016/j.fitote.2010.10.012Search in Google Scholar PubMed

46. Sa F, Gao J-L, Fung K-P, Zheng Y, Lee SM-Y, Wang Y-T. Anti-proliferative and proapoptotic effect of Smilax glabra Roxb. extract on hepatoma cell lines. Chem-Biol Interact 2008;171:1–14.10.1016/j.cbi.2007.08.012Search in Google Scholar PubMed

47. Xu S, Shang MY, Liu GX, Xu F, Wang X, Shou CC, et al. Chemical constituents from the rhizomes of Smilax glabra and their antimicrobial activity. Molecules 2013;18:5265–87.10.3390/molecules18055265Search in Google Scholar PubMed PubMed Central

48. Ao C, Higa T, Khanh TD, Upadhyay A, Tawata S. Antioxidant phenolic compounds from Smilax sebeana Miq. LWT Food Sci Technol 2011; 44:1681–6.10.1016/j.lwt.2011.02.001Search in Google Scholar

49. Wu LS, Wang XJ, Wang H, Yang HW, Jia AQ, Ding Q. Cytotoxic polyphenols against breast tumor cell in Smilax china L. J Ethnopharmacol 2010;130:460–4.10.1016/j.jep.2010.05.032Search in Google Scholar PubMed

50. Abdalaa S, Martin-Herrera D, Benjumea D, Gutierrez SD. Diuretic activity of some Smilax canariensis fractions. J Ethnopharmacol 2012;140:227–8.10.1016/j.jep.2012.01.017Search in Google Scholar PubMed

51. Huang AC, Wilde A, Ebmeyer J, Skouroumounis GK, Taylor DK. Examination of the phenolic profile and antioxidant activity of the leaves of the Australian native plant Smilax glyciphylla. J Nat Prod 2013;76:1930–6.10.1021/np4005163Search in Google Scholar PubMed

52. Li YL, Gan GP, Zhang HZ, Wu HZ, Li CL, Huang YP, et al. A flavonoid glycoside isolated from Smilax china L. rhizome in vitro anticancer effects on human cancer cell lines. J Ethnopharmacol 2007;113:115–24.10.1016/j.jep.2007.05.016Search in Google Scholar PubMed

53. Shao B, Guo HZ, Cui YJ, Liu AH, Yu HL, Guo H, et al. Simultaneous determination of six major stilbenes and flavonoids in Smilax china by high performance liquid chromatography. J Pharm Biomed Anal 2007;44:737–42.10.1016/j.jpba.2007.03.008Search in Google Scholar PubMed

54. Xu J, Feng S, Wang Q, Zhang M, Zhang C. A new flavonoid glycoside from the rhizomes and roots of Smilax scobinicaulis. Nat Prod Res 2014;28:517–21.10.1080/14786419.2014.880913Search in Google Scholar PubMed

55. Ozsoy N, Can A, Yanardag R, Akev N. Antioxidant activity of Smilax excelsa L. leaf extracts. Food Chem 2008;110:571–83.10.1016/j.foodchem.2008.02.037Search in Google Scholar

56. Seo HK, Lee JH, Kim HS, Lee CK, Lee SC. Antioxidant and antimicrobial activities of Smilax china L. leaf extracts. Food Sci Biotechnol 2012;21:1723–7.10.1007/s10068-012-0229-4Search in Google Scholar

Received: 2014-10-18

Accepted: 2015-3-20

Published Online: 2015-4-22

Published in Print: 2015-9-1

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https://doi.org/10.1515/jbcpp-2014-0114

Keywords for this article

antioxidant activity; brine shrimp; DPPH; H2O2; MCF7; MTT; Smilax zeylanica