PHYTOCHEMICAL ANALYSIS, ASSESSMENT OF ANTIPROLIFERATIVE AND FREE RADICAL SCAVENGING ACTIVITY OF MORUS ALBA AND MORUS RUBRA FRUITS


Farouk K El-baz, Amal Z Hassan, Howaida I Abd-alla, Hanan F Aly, Khaled Mahmoud

Abstract


Objective: Mulberry is a nontoxic commonly eaten plant, belongs to the Morus and used in folk medicine in the remedy of dysentery, antiphlogistic, diuretic, expectorant, and antidiabetic. The purpose of this study is to evaluate the antiproliferative and radical scavenging activity of the total alcoholic and successive fractions thereof of Morus alba and Morus rubra fruits. In addition, the chemical composition of the bioactive fractions of each species was investigated.

Methods: The antiproliferative potential of 8 extracts on 4 human cancer cell lines, hepatocellular carcinoma (HepG2), Caucasian breast adenocarcinoma (MCF7), prostate (PC3), and colon carcinoma (HCT116) in addition to one normal cell line namely human normal immortalized skin fibroblast cells (BJ1) were carried out. Cell viability was determined using MTT assay. The potency was compared with the reference drug doxorubicin. These extracts were also assayed for 1,1-diphenyl-2-hydrazyl free radical scavenging activities. After saponification of the n-hexane fraction, unsaponifiable matter and fatty acid methyl esters were analyzed by gas liquid chromatography (GLC). The chemical composition of the bioactive fractions was investigated using gas chromatography/mass spectrometry (GC/MS) analysis.

Results: All the extracts showed significant free radical scavenging activity dose-dependently. The n-hexane and dichloromethane (DCM) fractions of M. rubra exhibited potent cytotoxic activity on almost cancer cell lines. In the same pattern, ethyl acetate (EtOAc) of M. rubra has moderate cytotoxic activity against all cell lines except HepG2. DCM fraction of M. alba possessed both radical scavenging and high potential antiproliferated activities against HCT116 and MCF7 with inhibitory concentration of 43.9 and 32.3 μg/ml, respectively, while it showed no cytotoxic effect on BJ1. GLC analysis showed the major hydrocarbons in M. alba and M. rubra were heptacosane and docosane, respectively. Sterols were similar in both species but with different ratios and cholesterol was the major one. Palmitic and margaric were the major saturated fatty acid while arachidonic was the major unsaturated fatty acid in both species. GC/MS analysis showed the main compound in DCM fraction of each Morus species was palmitic acid. Furthermore, 1,11-bis-(methoxycarbonyl-ethenyl)-10,2-dihydroxy-cycloeicosane and linolelaidic acid, methyl ester were the main compounds in the EtOAc fraction of each Morus species. Whereas, the main compounds in alcoholic extract of M. alba and M. rubra were methyl-14-methyl-pentadecanoate and 1,2-O-isopropylyidene-4-nonene-1,2,3-triol, respectively.

Conclusions: The results observed remarkable biological activity of the successive fractions of M. rubra more than those of M. alba and confirmed its importance as a natural bioactive source. Morus species are good candidates to be promising as possible sources for future antitumor and antioxidants in food and pharmaceutical formulations. The strong activity partly explains the potential effects of Morus species for the treatment of cancer and degenerative diseases caused by free radicals.


Keywords


Morus alba, Morus rubra, Radical scavenging, Antiproliferated activity, BJ1, Gas liquid chromatography, Gas chromatography/mass spectrometry.

| PDF |

References


Cragg GM, Newman DJ. Natural products: A continuing source of novel drug leads. Biochim Biophys Acta 2013;1830(6):3670-95.

Lobo V, Patil A, Phatak A, Chandra N. Free radicals, antioxidants and functional foods: Impact on human health. Pharmacogn Rev 2010;4(8):118-26.

Awad HM, Abd-Alla HI, Mahmoud KH, El-Toumy SA. In vitro anti-nitrosative, antioxidant, and cytotoxicity activities of plant flavonoids: A comparative study. Med Chem Res 2014;23(7):3298-307.

Kähkönen MP, Heinämäki J, Ollilainen V, Heinonen M. Berry anthocyanins: Isolation, identification and antioxidant activities. J Sci Food Agric 2003;83(14):1403-11.

Burlando B, Clericuzio M, Cornara L. Moraceae plants with tyrosinase Inhibitory activity: A review. Mini Rev Med Chem 2017;17(2):108-21.

Ahmed HH, Hegazi MM, Abd-Alla HI, Eskander EF, Ellithey MS. Antitumour and antioxidant activity of some Red Sea seaweeds in ehrlich ascites carcinoma in vivo. Z Naturforsch C 2011;66(7-8):367-76.

Bishayee A, Politis T, Darvesh AS. Resveratrol in the chemoprevention and treatment of hepatocellular carcinoma. Cancer Treat Rev 2010;36(1):43-53. 8. Takuya K, Yukikazu Y. Apoptosis-inducing activity of ethanol extracts from the tea of mulberry (Morus alba) leaves in HL-60 cells. J Jpn Soc Food Sci Technol 2009;49(4):195-8.

Wei H, Zhu J, Liu XQ, Feng WH, Wang ZM, Yan LH. Review of bioactive compounds from root barks of Morus plants (Sang-Bai-Pi) and their pharmacological effects. Cogent Chem 2016;2. DOI: 10.1080/23312009.2016.1212320.

Aly HF, Abd-Alla HI, Ali SA, Mamdooh M, Alez RA, Abu-Krisha MT. Statistical data analysis which result from the bio-diagnosis and bio-treatment of injured rats with the hyperlipidemia and hyperglycemia diseases. Asian J Pharm Clin Res 2016;9(3):122-30.

Abd-Alla HI, Elnenaey HM, Hassan AZ, Taie HA, Abo-Shnaf RI, Hussein AM. Bioactive metabolites from two local cultivars of Ricinus communis and their free radical scavenging and acaricidal activities. Der Pharm Chem 2015;7(4):5-18.

Mosmann T. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immunol Methods 1983;65(1-2):55-63.

Thabrew MI, Hughes RD, McFarlane IG. Screening of hepatoprotective plant components using a HepG2 cell cytotoxicity assay. J Pharm Pharmacol 1997;49(11):1132-5.

Goveas S, Abraham A. Evaluation of antimicrobial and antioxidant activity of stem and leaf extracts of Coscinium fenestratum. Asian J Pharm Clin Res 2013;6(3):218-21.

Johnson AR, Davenport JB. Biochemistry and Methodology of Lipids. New York: John Wiley and Sons, Inc.; 1971. p. 31.

Vogel AI. Textbook of Practical Organic Chemistry. 3rd ed. London: Longman’s Green and Co., Ltd.; 1966. p. 133-6.

Adams RP. Identification of Essential Oils by Ion Trap Mass Spectroscopy. New York: Academic Press Inc.; 1995.

Jyoti A, Anand K, Sunanda P. Synergistic action of phytochemicals augments their antioxidative efficacy: An in vitro comparative study. Asian J Pharm Clin Res 2013;6(4):121-6.

Fathy SA, Singab AB, Nagwa SA, Abd El-Hamid DM, Zahra FA, Abd El-Moneim SM. The antiproliferative effect of mulberry (Morus alba L.) plant on hepatocarcinoma cell line HepG2. Egypt J Med Hum Genet 2013;14(4):375-82.

Ercisli S, Tosun M, Duralija B, Voca S, Sengul M, Turan M. Phytochemical content of some black (Mors nigra L.) and purple (Morus rubra L.) mulberry genotypes. Food Technol Biotechnol 2010;48(1):102-6.

Chen PN, Chu SC, Chiou HL, Kuo WH, Chiang CL, Hsieh YS. Mulberry anthocyanins, cyanidin 3-rutinoside and cyanidin 3-glucoside, exhibited an inhibitory effect on the migration and invasion of a human lung cancer cell line. Cancer Lett 2006;235(2):248-59.

El-Baz FK, Khalil WK, Aly HF, Booles HF. Berry extracts improved inflammatory cytokines, antioxidant enzyme and suppressed the gene expression alterations in diabetic rats. Int J Pharm Pharm Sci 2016;8:1-9.

Taie HA, Abd-Alla HI, Ali SA, Aly HF. Chemical composition and biological activities of two Solanum tuberosum cultivars grown in Egypt. Int J Pharm Pharm Sci 2015;7(6):311-20.

Naowaratwattana W, De-Eknamkul W, De Mejia EG. Phenolic-containing organic extracts of mulberry (Morus alba L.) leaves inhibit HepG2 hepatoma cells through G2/M phase arrest, induction of apoptosis, and inhibition of topoisomerase IIa activity. J Med Food 2010;13(5):1045-56.

Lin JY, Tang CY. Determination of total phenolics and flavonoid contents in selected fruits and vegetables, as well as their stimulatory effects on mouse splenocyte proliferation. Food Chem 2007;101(1):140-7.

Valko M, Leibfritz D, Moncol J, Cronin MT, Mazur M, Telser J. Free radicals and antioxidants in normal physiological functions and human disease. Int J Biochem Cell Biol 2007;39(1):44-84.

Gago-Dominguez M, Jiang X, Castelao JE. Lipid peroxidation, oxidative stress genes and dietary factors in breast cancer protection: A hypothesis. Breast Cancer Res 2007;9(1):201.

Hecht F, Pessoa CF, Gentile LB, Rosenthal D, Carvalho DP, Fortunato RS, et al. The role of oxidative stress on breast cancer development and therapy. Tumour Biol 2016;37(4):4281-91.

Hwang YJ, Lee EJ, Kim HR, Hwang KA. In vitro antioxidant and anticancer effects of solvent fractions from Prunella vulgaris var. Lilacina. BMC Complement Altern Med 2013;13:310.

Menéndez JA, del Mar Barbacid M, Montero S, Sevilla E, Escrich E, Solanas M, et al. Effects of gamma-linolenic acid and oleic acid on paclitaxel cytotoxicity in human breast cancer cells. Eur J Cancer 2001;37(3):402-13.

Achenef B, Arifah K. Cytotoxic effects of conjugated linoleic acids on human breast cancer cells (MCF7). Acad J Cancer Res 2012;5(1):11-6.

Harada H, Yamashita U, Kurihara H, Fukushi E, Kawabata J, Kamei Y. Antitumor activity of palmitic acid found as a selective cytotoxic substance in a marine red alga. Anticancer Res 2002;22(5):2587-90.

Mbimba T, Awale P, Bhatia D, Geldenhuys WJ, Darvesh AS, Carroll RT, et al. Alteration of hepatic proinflammatory cytokines is involved in the resveratrol-mediated chemoprevention of chemically-induced hepatocarcinogenesis. Curr Pharm Biotechnol 2012;13(1):229-34.

Figueiredo CR, Matsuo AL, Pereira FV, Rabaça AN, Farias CF, Girola N, et al. Pyrostegia venusta heptane extract containing saturated aliphatic hydrocarbons induces apoptosis on B16F10-Nex2 melanoma cells and displays antitumor activity in vivo. Pharmacogn Mag 2014;10 Suppl 2:S363-76.

Yashvanth S, Rani SS, Madhavendra SS. Morus alba L. A new perspective: Scanning electron microscopic, micro chemical, GC-MS and UPLC-MS Characterisation. Int J Res Pharm Chem 2015;5(1):106-15.

de Azeredo SO, Figueroa-Villar JD. Phenanthrene derivatives for synthesis and applications in medicinal chemistry: A review. World J Pharm Pharm Sci 2014;3:1362-79.

Lima LA, Alves TM, Zani CL, Pimenta LP, Boaventura MA. Antioxidant and cytotoxic potential of fatty acid methyl esters from the seeds of Annona cornifolia A. St.-Hil. (Annonaceae). Food Res Int 2012;48(2):873-5.

Sayegh F, Elazzazy A, Bellou S, Moustogianni A, Elkady AI, Baeshen MN, et al. Production of polyunsaturated single cell oils possessing antimicrobial and anticancer properties. Ann Microb 2016;66(3):937-48.

Dimitrijević DS, Kostić DA, Stojanović GS, Mitić SS, Mitić MN, Đorđević AS, et al. A survey on macro - And micro-elements, phenolic compounds, biological activity and use of Morus spp. (Moraceae). Fruits 2013;68:333-47.




About this article

Title

PHYTOCHEMICAL ANALYSIS, ASSESSMENT OF ANTIPROLIFERATIVE AND FREE RADICAL SCAVENGING ACTIVITY OF MORUS ALBA AND MORUS RUBRA FRUITS

Keywords

Morus alba, Morus rubra, Radical scavenging, Antiproliferated activity, BJ1, Gas liquid chromatography, Gas chromatography/mass spectrometry.

DOI

10.22159/ajpcr.2017.v10i6.18029

Date

01-06-2017

Additional Links

Manuscript Submission

Journal

Asian Journal of Pharmaceutical and Clinical Research
Vol 10 Issue 6 June 2017 Page: 189-199

Print ISSN

0974-2441

Online ISSN

2455-3891

Statistics

52 Views | 20 Downloads

Authors & Affiliations

Farouk K El-baz
Department of Plant Biochemistry, Division of Agriculture and Biology Research, National Research Centre, Dokki-Giza 12622, Egypt
Egypt

Amal Z Hassan
Department of Chemistry of Natural Compounds, Division of Pharmaceutical Industries Research, National Research Centre, Dokki-Giza 12622, Egypt.
Egypt

Howaida I Abd-alla
Department of Chemistry of Natural Compounds, Division of Pharmaceutical Industries Research, National Research Centre, Dokki-Giza 12622, Egypt.
Egypt

Hanan F Aly
Department of Therapeutic Chemistry, Division of Pharmaceutical Industries, National Research Centre, Dokki-Giza 12622, Egypt.
Egypt

Khaled Mahmoud
Department of Pharmacognosy, Division of Pharmaceutical Industries, National Research Centre, Dokki-Giza 12622, Egypt.
Egypt


Article Tools


Email this article (Login required)
Email the author (Login required)

Refbacks

  • There are currently no refbacks.