Evaluation of the Genotoxic and Antigenotoxic Effects of Chios Mastic Water by the In Vitro Micronucleus Test on Human Lymphocytes and the In Vivo Wing Somatic Test on Drosophila
Chios mastic gum, a plant-derived product obtained by the Mediterranean bush Pistacia lentiscus (L.) var. chia (Duham), has generated considerable interest because of its antimicrobial, anticancer, antioxidant and other beneficial properties. Its aqueous extract, called Chios mastic water (CMW), contains the authentic mastic scent and all the water soluble components of mastic. In the present study, the potential genotoxic activity of CMW, as well as its antigenotoxic properties against the mutagenic agent mitomycin-C (MMC), was evaluated by employing the in vitro Cytokinesis Block MicroNucleus (CBMN) assay and the in vivo Somatic Mutation And Recombination Test (SMART). In the former assay, lymphocytes were treated with 1, 2 and 5% (v/v) of CMW with or without MMC at concentrations 0.05 and 0.50 μg/ml. No significant micronucleus induction was observed by CMW, while co-treatment with MMC led to a decrease of the MMC-induced micronuclei, which ranged between 22.8 and 44.7%. For SMART, larvae were treated with 50 and 100% (v/v) CMW with or without MMC at concentrations 1.00, 2.50 and 5.00 μg/ml. It was shown that CMW alone did not modify the spontaneous frequencies of spots indicating lack of genotoxic activity. Τhe simultaneous administration of MMC with 100% CMW led to considerable alterations of the frequencies of MMC-induced wing spots with the total mutant clones showing reduction between 53.5 and 74.4%. Our data clearly show a protective role of CMW against the MMC-induced genotoxicity and further research on the beneficial properties of this product is suggested.
References
[1]
Mills JS, White R (1977) Natural resins of art and archaeology: their sources, chemistry and identification. Stud Conserv 22: 12–31.
[2]
Margaris NS (1981) Adaptative strategies in plants dominating Mediterranean-type ecosystems. In: di Castri R, Goodall DW, Specht RI, editors. Ecosystems of the world, Mediterranean type Shrublands. New York: Elsevier Science. 309–315.
[3]
Serpico M (2000) Resins, amber and bitumen in ancient Egyptian materials and technology. Cambridge: Cambridge University Press. 430 p.
[4]
Doukas C (2003) Cosmetics that contain mastic gum and mastic oil. Chem Chron 12: 36–39.
[5]
Triantafyllou A, Bikineyeva A, Dikalova A, Nazarewicz R, Lerakis S, et al. (2011) Anti-inflammatory activity of Chios mastic gum is associated with inhibition of TNF-alpha induced oxidative stress. Nutr J 10: 64.
[6]
Tassou CC, Nychas GCE (1995) Antimicrobial activity of the essential oil of mastic gum (P. lentiscus var. chia) on Gram Positive and Gram Negative bacteria in broth and in model food system. Int Biodeter Biodegr 36: 411–420.
[7]
Iauk L, Ragusa A, Rapisarda A, Franco S, Nicolosi V (1996) In vitro antimicrobial activity of Pistacia lentiscus L. extracts: preliminary report. J Chemother 8: 207–209.
[8]
Magiatis P, Melliou E, Skaltsounis A, Chinou I, Mitaku S (1999) Chemical composition and antimicrobial activity of the essential oils of Pistacia lentiscus var. chia. Planta Med 65: 749–752.
[9]
Assimopoulou AN, Zlatanos SN, Papageorgiou VP (2005) Antioxidant activity of natural resins and bioactive triterpenes in oil substrates. Food Chem 92: 721–727.
[10]
Koutsoudaki C, Krsek M, Rodger A (2005) Chemical composition and antibacterial activity of the essential oil and the gum of Pistacia lentiscus var. chia. J Agric Food Chem 53: 7681–7685.
[11]
Zhou L, Satoh K, Takahashi K, Watanabe S, Nakamura W, et al. (2009) Reevaluation of anti-inflammatory activity of mastic using activated macrophages. In Vivo 23: 583–589.
[12]
Mahmoudi M, Ebrahimzadeh MA, Nabavi SF, Hafezi S, Nabavi SM, et al. (2010) Antiinflammatory and antioxidant activities of gum mastic. Eur Rev Med Pharmacol Sci 14: 765–769.
[13]
Paraschos S, Mitakou S (2012) Skaltsounis (2012) AL Chios gum mastic: a review of its biological activities. Curr Med Chem 19: 2292–2302.
[14]
Al-Habbal MJ, Al-Habbal Z, Huwez F (1984) A double-blind controlled clinical trial of mastic and placebo in the treatment of duodenal ulcer. J Clin Exp Pharmacol Physiol 11: 541–544.
[15]
Al-Said M, Ageel AM, Parmar NS, Tariq M (1986) Evaluation of mastic, a crude drug obtained from Pistacia lentiscus for gastric and duodenal anti-ulcer activity. J Ethnopharmacol 15: 271–278.
[16]
Takahashi K, Fukazawa M, Motohia H, Ochiai K, Nishikawa H, et al. (2003) A pilot study on antiplaque effects of mastic chewing gum in the oral cavity. J Periodontol 74: 501–505.
[17]
Dedoussis GV, Kaliora AC, Psarras S, Chiou A, Mylona A, et al. (2004) Antiatherogenic effect of Pistacia lentiscus via GSH restoration and downregulation of CD36 mRNA expression. Atherosclerosis 174: 293–303.
[18]
Kaliora AC, Stathopoulou MG, Triantafillidis JK, Dedoussis GV, Andrikopoulos NK (2007) Alterations in the function of circulating mononuclear cells derived from patients with Crohn’s disease treated with mastic. World J Gastroenterol 7: 6031–6036.
[19]
Triantafyllou A, Chaviaras N, Sergentanis TN, Protopapa E, Tsaknis J (2007) Chios mastic gum modulates serum biochemical parameters in a human population. J Ethnopharmacol 111: 43–49.
[20]
Dabos KJ, Sfika E, Blatta LJ, Frantzi D, Amygdalos GI, et al. (2010) Is Chios mastic gum effective in the treatment of functional dyspepsia? A prospective randomised double-blind placebo controlled trial. J Ethnopharmacol 127: 205–209.
[21]
Vallianou I, Peroulis N, Pantazis P, Hatzopoulou-Cladaras M (2011) Camphene, a plant-derived monoterpene, reduces plasma cholesterol and triglycerides in hyperlipidemic rats independently of HMG-CoA reductase activity. Plos One 6: 1–11.
[22]
Loutrari H, Magkouta S, Pyriochou A, Koika V, Kolisis FN, et al. (2006) Mastic oil from Pistacia lentiscus var. chia inhibits growth and survival of human K562 leukemia cells and attenuates angiogenesis. Nutr Cancer 55: 86–93.
[23]
He ML, Yuan HK, Jiang AL, Gong AY, Chen WW, et al. (2006) Gum mastic inhibits the expression and function of the androgen receptor in prostate cancer cells. Cancer 106: 2547–2555.
[24]
He ML, Li A, Xu CS, Wang SL, Zhang MJ, et al. (2007) Mechanisms of antiprostate cancer by gum mastic: NF-kappaB signal as target. Acta Pharmacol Sin 28: 446–452.
[25]
Balan KV, Prince J, Han Z, Dimas K, Cladaras M, et al. (2007) Antiproliferative activity and induction of apoptosis in human colon cancer cells treated in vitro with constituents of a product derived from Pistacia lentiscus L. var. chia. Phytomedicine 14: 263–272.
[26]
Magkouta S, Stathopoulos GT, Psallidas I, Papapetropoulos A, Kolisis FN, et al. (2009) Protective effects of mastic oil from Pistacia lentiscus variation chia against experimental growth of lewis lung carcinoma. Nutr Cancer 61: 640–648.
[27]
Huang XY, Wang HC, Yuan Z, Li A, He ML, et al. (2010) Gemcitabine combined with gum mastic causes potent growth inhibition and apoptosis of pancreatic cancer cells. Acta Pharmacol Sin 31: 741–745.
[28]
Giaginis C, Theocharis S (2011) Current evidence on the anticancer potential of Chios mastic gum. Nutr Cancer 63: 1174–1184.
[29]
Paraschos S, Magiatis P, Gousia P, Economou V, Sakkas H, et al. (2011) Chemical investigation and antimicrobial properties of mastic water and its major constituents. Food Chem 129: 907–911.
[30]
OECD website. Available: http://www.oecd.org/env/ehs/testing/TG48？7%20Oct%202012%20updated%2029oct.pdf. Accessed 2013 May 31.
[31]
Kirsch-Volders M, Decordier I, Elhajouji A, Plas G, Aardema MJ, et al. (2011) In vitro genotoxicity testing using the micronucleus assay in cell lines, human lymphocytes and 3D human skin models. Mutagenesis 26: 177–184.
[32]
Graf U, Würgler FE, Katz AJ, Frei H, Juon H, et al. (1984) Somatic mutation and recombination test in Drosophila melanogaster. Environ Mutagen 6: 153–188.
[33]
Graf U, Abraham SK, Guzman-Rincon J, Würler FE (1998) Antigenotoxicity studies in Drosophila melanogaster. Mutat Res 402: 203–209.
[34]
Banfi S, Borsani G, Rossi E, Bernard L, Guffanti A, et al. (1996) Identification and mapping of human cDNAs homologous to Drosophila mutant genes through EST database searching. Nat Genet 13: 167–174.
[35]
Reiter LT, Potocki L, Chien S, Gribskov M, Bier E (2001) A Systematic analysis of human disease-associated gene sequences in Drosophila melanogaster. Genome Res 11: 1114–1125.
[36]
Apidianakis Y, Rahme LG (2011) Drosophila melanogaster as a model for human intestinal infection and pathology. Dis Model Mech 4: 21–30.
[37]
Kim SI, Jung JW, Ahn YJ, Restifo LL, Kwon HK (2011) Drosophila as a model system for studying lifespan and neuroprotective activities of plant-derived compounds. J Asia-Pac Entomol 14: 509–517.
[38]
Kounatidis I, Ligoxygakis P (2012) Drosophila as a model system to unravel the layers of innate immunity to infection. Open Biol 2: 120075.
[39]
Bonassi S, El-Zein R, Bolognesi C, Fenech M (2011) Micronuclei frequency in peripheral blood lymphocytes and cancer risk: evidence from human studies. Mutagenesis 26: 93–100.
[40]
Sengstag C (1994) The role of mitotic recombination in carcinogenesis. Crit Rev Toxicol 24: 323–353.
[41]
Clare G, Lorenzon G, Akhurst L, Marzin D, van Delft J, et al. (2006) SFTG International collaborative study on the in vitro micronucleus test. II. Using human lymphocytes. Mutat Res 607: 37–60.
[42]
Surrallés J, Carbonell E, Marcos R, Degrassi F, Antoccia A, et al. (1992) A collaborative study on the improvement of the micronucleus test in cultured human lymphocytes. Mutagenesis 7: 407–410.
[43]
Vlastos D, Stephanou G (1998) Effects of cetirizine dihydrochloride on human lymphocytes in vitro: micronucleus induction. Evaluation of clastogenic and aneugenic potential using CREST and FISH assays. Arch Dermatol Res 290: 312–318.
[44]
Papapaulou P, Vlastos D, Stephanou G, Demopoulos NA (2001) Linuron cytogenetic activity on human lymphocutes treated in vitro. Evaluation of clastogenic and aneugenic potential using Cytokinesis Block Micronucleus Assay in combination with Fluorescence in situ Hybridization (FISH). Fresen Environ Bull 10: 421–437.
[45]
Demsia G, Vlastos D, Goumenou M, Matthopoulos DP (2007) Assessment of the genotoxicity of imidacloprid and metalaxyl in cultured human lymphocytes and rat bone marrow. Mutat Res 634: 32–39.
[46]
Fenech M (1997) The advantages and disadvantages of the cytokinesis-block micronucleus method. Mutat Res 392: 11–18.
[47]
Fenech M, Chang WP, Kirsch-Volders M, Holland N, Bonassi S, et al. (2003) HUMN project: detailed description of the scoring criteria for the cytokinesis-block micronucleus assay using isolated human lymphocyte cultures. Mutat Res 534: 65–75.
[48]
Surrallés J, Xamena N, Creus A, Catalan J, Norppa H, et al. (1995) Induction of micronuclei by five pyrethroid insecticides in whole-blood and isolated human lymphocyte cultures. Mutat Res 341: 169–184.
[49]
Lindsley DL, Zimm GG (1992) The genome of Drosophila melanogaster. San Diego: Academic Press. 1133 p.
[50]
Marec F, Gelbic I (1994) High recombinagenic activities of three antiviral agents adenine derivatives, in the Drosophila wing spot test. Mutat Res 311: 305–317.
[51]
Graf U, van Schaik N (1992) Improved high bioactivation cross for the wing somatic mutation and recombination test in Drosophila melanogaster. Mutat Res 271: 59–67.
[52]
Selby PB, Olson WH (1981) Methods and criteria for deciding whether specific- locus mutation-rate data in mice indicate a positive, negative, or inconclusive result. Mutat Res 83: 403–418.
[53]
Frei H, Würgler FE (1988) Statistical methods to decide whether mutagenicity test data from Drosophila assays indicate a positive, negative, or inconclusive result. Mutat Res 203: 297–308.
[54]
Kastenbaum MA, Bowman KO (1970) Tables for determining the statistical significance of mutation frequencies. Mutat Res 9: 527–549.
[55]
Margolin BH, Collings BJ, Mason JM (1983) Statistical analysis and sample-size determinations for mutagenicity experiments with binomial responses. Environ Mutagen 5: 705–716.
[56]
Frei H, Würgler FE (1995) Optimal experimental design and sample size for the statistical evaluation of data from somatic mutation and recombination tests (SMART) in Drosophila. Mutat Res 334: 247–258.
[57]
Abraham SK (1994) Antigenotoxicity of coffee in the Drosophila assay for somatic mutation and recombination. Mutagenesis 9: 383–386.
[58]
Kang JS, Wanibuchi H, Salim EI, Kinoshita A, Fukushima S (2007) Evaluation of the toxicity of mastic gum with 13 weeks dietary administration to F344 rats. Food Chem Toxicol 45: 494–501.
[59]
Loizzo MR, Tundis R, Menichini F, Saab AM, Statti GA, et al. (2008) Antiproliferative effects of essential oils and their major constituents in human renal adenocarcinoma and amelanotic melanoma cells. Cell Proliferat 41: 1002–1012.
[60]
Garozzo A, Timpanaro R, Bisignano B, Furneri PM, Bisignano G, et al. (2009) In vitro antiviral activity of Melaleuca alternifolia essential oil. Lett Appl Microbiol 49: 806–808.
[61]
Hortháthová E, Slameňová D, Mar？álková L, ？ramková M, Wsólová L (2009) Effects of borneol on the level of DNA damage induced in primary rat hepatocytes and testicular cells by hydrogen peroxide. Food Chem Toxicol 47: 1318–1323.
[62]
Kim HJ, Neophytou K (2009) Natural anti-Inflammatory compounds for the management and adjuvant therapy of inflammatory bowel disease and its drug delivery system. Arch Pharm Res 32: 997–1004.
[63]
Slameňová D, Hortháthová E, Wsólová L, ？ramková M, Navarová J (2009) Investigation of anti-oxidative, cytotoxic, DNA-damaging and DNA-protective effects of plant volatiles eugenol and borneol in human-derived HepG2, Caco-2 and VH10 cell lines. Mutat Res 677: 46–52.
[64]
Nibret E, Wink M (2010) Trypanocidal and antileukaemic effects of the essential oils of Hagenia abyssinica, Leonotis ocymifolia, Moringa stenopetala, and their main individual constituents. Phytomedicine 17: 911–920.
[65]
Mademtzoglou D, Akmoutsou P, Kounatidis I, Franzios G, Drosopoulou E, et al. (2011) Applying the Drosophila wing spot test to assess the genotoxic impact of 10 essential oil constituents used as flavouring agents or cosmetic ingredients. Flavour Fragr J 26: 447–451.
[66]
Bestmann HJ, Classen B, Vostrowsky O, Klingauf F, Stein U (1988) Botanical insecticides, VIII: the synergistic effect of carvone and pyrethrin I in the essential oil of Chrysanthemum balsamita L. J Appl Entomol. 106: 144–149.
[67]
Franzios G, Mirotsou M, Hatziapostolou E, Kral J, Scouras ZG, et al. (1997) Insecticidal and genotoxic activities of mint essential oils. J Agric Food Chem 45: 2690–2694.
[68]
Abdelwahed A, Bouhlel I, Skandrani I, Valenti K, Kadri M, et al. (2007) Study of antimutagenic and antioxidant activities of gallic acid and 1,2,3,4,6-pentagalloylglucose from Pistacia lentiscus. Confirmation by microarray expression profiling. Chem Biol Interact 165: 1–13.
[69]
Abdelwahed A, Bhouri W, Neffati A, Ben Sghaier M, Boubaker J, et al. (2009) Antigenotoxic and antioxidant activities of fruit extracts from (Tunisian) Pistacia lentiscus. Food Sci Tech Int 15: 215–222.
[70]
Abraham SK, Singh SP (1999) Anti-genotoxicity and glutathione S-transferase activity in mice pretreated with caffeinated and decaffeinated coffee. Food Chem Toxicol 37: 733–739.
[71]
Lehmann M, Graf U, Reguly ML, Rodrigues De Andrade HH (2000) Interference of tannic acid on the genotoxicity of mitomycin C, methylmethanesulfonate, and nitrogen mustard in somatic cells of Drosophila melanogaster. Environ Mol Mutagen 36: 195–200.
[72]
Niikawa M, Shin S, Nagase H (2007) Suppressive effect of post- or pre-treatment of aspirin metabolite on mitomycin C-induced genotoxicity using the somatic mutation and recombination test in Drosophila melanogaster. Biomed Pharmacother 61: 113–119.
[73]
Speit G, Linsenmeyer R, Schütz P, Kuehner S (2012) Insensitivity of the in vitro cytokinesis-block micronucleus assay with human lymphocytes for the detection of DNA damage present at the start of the cell culture. Mutagenesis 27: 743–747.
[74]
Turkez H, Aydin E, Aslan A (2012) Xanthoria elegans (Link) (lichen) extract counteracts DNA damage and oxidative stress of mitomycin C in human lymphocytes. Cytotechnology 64: 679–686.
[75]
Lorge E, Thybaud V, Aardema MJ, Oliver J, Wakata A, et al. (2006) SFTG International collaborative study on the in vitro micronucleus test. I. General conditions and overall conclusions of the study. Mutat Res 607: 13–36.
[76]
Al-Zubairi AS, Abdul AB, Syam MM (2010) Evaluation of the genotoxicity of zerumbone in cultured human peripheral blood lymphocytes. Toxicol In Vitro 24: 707–712.
[77]
Mazumdar M, Giri S, Giri A (2011) Role of quercetin on mitomycin C induced genotoxicity: analysis of micronucleus and chromosome aberrations in vivo. Mutat Res 721: 147–152.
[78]
Doi K, Wei M, Kitano M, Uematsu N, Inoue M, et al. (2009) Enhancement of preneoplastic lesion yield by Chios Mastic Gum in a rat liver medium-term carcinogenesis bioassay. Toxicol Appl Pharm 234: 135–142.
[79]
Kaya B, Creus A, Velázquez A, Yanikoglu A, Marcos R (2002) Genotoxicity is modulated by ascorbic acid. Studies using the wing spot test in Drosophila. Mutat Res 520: 93–101.
[80]
De Rezende AAA, e Silva MLA, Tavares DC, Cunha WR, Rezende KCS, et al. (2011) The effect of the dibenzylbutyrolactolic lignan (-)-cubebin on doxorubicin mutagenicity and recombinogenicity in wing somatic cells of Drosophila melanogaster. Food Chem Toxicol 49: 1235–1241.
[81]
Resende FA, Tomazella IM, Barbosa LC, Ponce M, Furtado RA, et al. (2011) Effect of the dibenzylbutyrolactone lignan (？)-hinokinin on doxorubicin and methyl methanesulfonate clastogenicity in V79 Chinese hamster lung fibroblasts. Mutat Res 700: 62–66.
[82]
De Rezende AAA, Munari CC, de Oliveira PF, Ferreira NH, Tavares DC, et al. (2013) A comparative study of the modulatory effects of (-)-cubebin on the mutagenicity/recombinogenicity induced by different chemical agents. Food Chem Toxicol 55: 645–652.
[83]
Kato FH, Viana NI, Santini CB, de Souza CG, Veneziani RC, et al. (2012) Assessment of the in vitro and in vivo genotoxic and antigenotoxic effects of pimaradienoic acid in mammalian cells. Mutat Res 749: 87–92.
[84]
Miti？-？ulafi？ D, ？egura B, Nikoli？ B, Vukovi？-Ga？i？ B, Kne？evi？-Vuk？evi？ J, et al. (2009) Protective effect of linalool, myrcene and eucalyptol against t-butyl hydroperoxide induced genotoxicity in bacteria and cultured human cells. Food Chem Toxicol 47: 260–266.
[85]
Chan NLS, Wang H, Wang Y, Leung HY, Leung LK (2006) Polycyclic aromatic hydrocarbon-induced CYP1B1 activity is suppressed by perillyl alcohol in MCF-7 cells. Toxicol Appl Pharm 213: 98–104.
[86]
De Flora S, Ramel C (1988) Mechanisms of inhibitors of mutagenesis and carcinogénesis. Classification and overview. Mutat Res 202: 285–306.
[87]
Hodnick WF, Sartorelli AC (1993) Reductive activation of mitomycin C by NADH: cytochrome b5 reductase. Cancer Res 53: 4907–4491.
[88]
Tomasz M, Palom Y (1997) The mitomycin bioreductive antitumor agents: cross-linking and alkylation of DNA as the molecular basis of their activity. Pharmacol Ther 76: 73–87.
[89]
Bakkali F, Averbeck S, Averbeck D, Idaomar M (2008) Biological effects of essential oils–A review. Food Chem Toxicol 46: 446–475.
[90]
De Flora S, Izzotti A (2007) Mutagenesis and cardiovascular diseases: molecular mechanisms, risk factors, and protective factors. Mutat Res 621: 5–17.
[91]
Jones S, Chen WD, Parmigiani G, Diehl F, Beerenwinkel N, et al. (2008) Comparative lesion sequencing provides insight into tumor evolution. Proc Natl Acad Sci USA 105: 4283–4288.
[92]
Erickson RP (2010) Somatic gene mutation and human disease other than cancer: an update. Mutat Res 705: 96–106.
[93]
Frank SA (2010) Evolution in health and medicine Sackler colloquium: Somatic evolutionary genomics: mutations during development cause highly variable genetic mosaicism with risk of cancer and neurodegeneration. Proc Natl Acad Sci USA 107 Suppl 11725–1730.
