AYFER BECEREN, Betül SARIKAYA, MÜCTEBA EŞREF TATLIPINAR, Gülden Z. OMURTAG, SEMRA ŞARDAŞ

Antioksidan Maddelerin Mutajenite Üzerine Etkilerinin Ames Testi ile İncelenmesi

Günümüzde insan sağlığında önemli rol oynayan kimyasal ajanların mutajenik etkilerinin gözlem altına alınmasını sağlayan çeşitli test sistemleri geliştirilmiştir. Bu sistemlerden biri de Ames testidir. Ames testi ile kimyasalların mutajenik etkilerinin belirlenmesi için çeşitli mutant bakteriler geliştirilmiştir. Bunun için de Salmonella typhimurium bakterisinin çeşitli suşları kullanılmaktadır. Bu çalışmada Salmonella/mikrozom Ames test kiti ile metabolik aktivasyon varlığında ve yokluğunda, gen mutasyonlarına sebep olan karsinojenik madde olan 2-Aminoflorene karşı antioksidan etkiye sahip Pelargonium sidoides’in muhtemel antimutajenik etkisinin araştırılması amaçlanmıştır. Yöntemde Salmonella typhimurium TA 100 ve TA 98 suşları ile çalışılmıştır. TA 100 baz çifti değişimine, TA 98 ise çerçeve kaymasına yol açan mutajenlerin belirlenmesi için kullanılmıştır. Deneyler S9’lu ve S9’suz olmak üzere iki grup halinde gerçekleştirilmiştir. Test sonuçlarının ortalaması alınarak, pozitif ve negatif kontrol grupları ile karşılaştırılmış ve değerlendirilmiştir. Çalışmamızda, Pelargonium sidoides’in TA 100 ve TA 98 suşları ile S9 yokluğunda 2-aminofluoren’e karşı antimutajenik aktiviteye sahip olduğu (p≥0.05) S9 varlığında ise antimutajenik aktiviteye sahip olmadığı (p≤0.05) gösterilmiştir.

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Barile FA. Principle of Toxicology testing page, Chapter 15. New York, USA. 2008, pp 209-228.

Gatehouse D, Haworth S, Cebula T, Gocke E, Kier L, Matsushima T, Melcion C, Nohmi T, Ohta T, Venitt S, Zeiger E. Recommendations for the performance of bacterial mutation assays. Mutat Res 1994;312: 217-33.

Kier LE, Brusick DJ, Auletta AE, Von Halle ES, Brown MM, Simmon VF, Dunkel V, McCann J, Mortel-mans K, Prival M, Rao TK, Ray V. The Salmonella typhimurium mammalian microsomal assay: A report of the U.S. Environmental Protection Agency Gene-Tox Program. Mutat Res 1986;168: 69–240.

Pai CA. Foundation of genetics. Asciense for society. Keffort press, Singapur. 1985.

Zeiger E, Anderson B, Haworth S, Lawlor T, Mortelmans K, Salmonella mutagenicity tests V. Results from the testing of 311 chemicals. Environ Mol Mutagen 1992;19: 2–141.

Temizkan GO. Genetik. İstanbul Universitesi Fen Fakultesi Basımevi, İstanbul. 1994, s: 281.

Mortelmans K, Zeiger E. The Ames Salmonella/microsome mutagenicity assay. Mutat Res 2000;455: 29-60.

Ames BN, McCann J,Yamasaki E. Methods for detecting carcinogens and mutagens with the Salmonella/mammalianmicrosome mutagenicity test. Mutat Res 1975;31:347-64.

Regulatory Genetic Toxicology Tests. In: Genetic Toxicology and Cancer Risk Assessment. Editor: Choy WN. New York. 2001, pp 93-114.

Maron DM, Ames BN. Revised methods for the mutagenicity test. Mutat Res 1983;113: 173-215.

Umbuzeiro Gde A, Rech CM, Correia S, Bergamasco AM, Cardenette GH, Fluckiger-Isler S, Kamber M. Comparison of the Salmonella/microsome microsuspension assay with the new microplate fluctuation protocol for testing the mutagenicity of environmental samples. Environ Mol Mutagen 2010;51: 31-8.

Kado NY, Langley D, Eisenstadt E. A simple modification of the Salmonella liquid incubation assay. Mutat Res 1983;121: 25-32.

Fluckiger-Isler S, Baumeister M, Braun K, Gervais V, Hasler- Nguyen N, Reimann R, Van Gompel J, Wunderlich HG, Engelhardt G. Assessment of the performance of the Ames II assay: a collaborative study with 19 coded compounds. Mutat Res 2004;558: 181-97.

Gee P, CH. Sommers, A. S. Melick, X. M. Gidrol, M. D. Todd, R. B. Burris, M. E. Nelson & E. Zeiger. Comparison of responses of base-specific Salmonella tester strains with the traditional strains for identifying mutagens: the results of a validation study. Mutat Res 1998;412: 115-30.

Green MHL, Muriel WJ, Bridges BA. Use of a simplified fluctuation test to detect low levels of mutagens. Mutat Res 1976;38: 33–42.

Sghaiera MB, Boubakera J, Skandrani I, Bouhlel I, Limema I, Ghediraa K, Chekir-Ghedira L. Antimutagenic, antigenotoxic and antioxidant activities of phenolic-enriched extracts from Teucrium ramosissimum: Combination with their phytochemical composition. Environ Toxicol Pharmacol 2010;31: 220-32.

Zegura B, Dobnik D, Niderl MH, Flipic M. Antioxidant and antigenotoxic effects of rosemary (Rosmarinus officinalis L.) extracts in Salmonella typhimurium TA98 and HepG2 cells. Environ Toxiol Pharmacol 2011;32: 296-305.

Gervais V, Bijot D, Claude N. Assessment of a screening experience with the Ames II test and future prospects. (servier) European Environmental Mutagen Society Annual Meeting: From Hazard to risk, Aberdeen Scotland (UK). 2003, pp 120.

Kamber M, Fluckiger-Isler S, Engelhardt G, Jaeckkh R, Zeigerr E. Comparison of the Ames II and traditional Ames test responses with respect to mutagenicity, strain specificities, need for metabolism and correlation with rodent carcinogenicity. Mutagenesis 2009;24: 359-66.

Kolodziej H, Kiderlen AF. İn vitro evaluation of antibacterial and immunomodulatory activities of Pelargonium reniforme, Pelargonium sidoides and the releted herbal drug preparation EPs 7630. Phytomed 2007;14: 18-26.

Ancos B, Gonzalez EM, Cano MP. Ellagic Acid, Vitamin C and total phenolic contents and radical scavenging capacity affected by freezingand frozen storage in raspberry fruit. J Agrıc Food Chem 2000;48: 4565-70.

Beudot C, De Me´o MP, Dauzonne D, Eliasc R, Laget M, Guiraud H, Balansard G, Dume´nil G. Evaluation of the mutagenicity and antimutagenicity of forty-two 3-substituted flavones in the Ames Test. Mutat Res 1998;417: 141-53.

Chai Y, Luo Q, Sun M, Corke H. Antioxidant Activity and Phenolic Compounds of 112 Traditional Chinese Medicinal Plants Associated with Anticancer. Life Sci 2004;74: 2157-84.

Hayder N, Skandrani I, Kilani S, Bouhlel I, Abdelwahed A, Ben Ammar R, Mahmoud A, Ghedira K, Ghedira L. Antimutagenic activity of Myrtus communis L. using the Salmonella microsome assay. S Afr J Bot 2008;74: 121–5.

Ozbek T, Gulluce M, Şahin F, Sevsay S, Barış O. Investigation of the antimutagenic potentials of the methanol extract of Origanum vulgare L. Subsp. vulgare in the Eastern Anotolia region of Turkey. Turk J Biol 2008; 32: 271-6.

Ham S, Kim SH, Moon SY, Chung MJ, Cui CB, Han EK, Chung CK, Choe M. Antimutagenic effects of subfractions of Chaga mushroom(Inonotus obliquus) Extract. Mutat Res 2009;672: 55–9.

Sotto AD, Vitalone A, Nicoletti M, Piccin A, Mazzanti G. Pharmacological and phytochemical study on a Sisymbrium officinale Scop. Extract. J Ethnopharmacol 2010;127: 731–6.

Santhosh KT, Swarnam J, Ramadasan K. Potent suppressive effect of gren tea polyphenols on tobacco-induced mutagenicity. Phytomed 2005;12: 216-20.

İpek E, Zeytinoglu H, Okay S, Tuylu B, Kurkcuoglu M, Can Baser KH. Genotoxicity and antigenotoxicity of Origanum oil and carvacrol evaluated by Ames Salmonella/microsomal test. Food Chem 2005;93: 551-6.

Kaur S, Arora S, Kaur K, Kumar S. The in vitro antimutagenic activity of Triphala- an Indian herbal drug. Food Chem Toxicol 2002;40: 527-34.

Birt DF, Walker B, Tıbbels MG, Bresnick E. Antimutagenesis and anti-promotion by apigenin, robinetin and indole-3- carbinol. Carcinogenesis 1986;7: 959-63.

Raucher R, Edenharder R, Platt KL. In vitro antimutagenic and in vivo anticlastogenic effects of carotenoids and solvent extracts from fruits and vegetables rich in carotenoids. Mutat Res 1998;413: 129- 42.

Mativandlela SPN, Lall N, Meyer JJM. Antibacterial, antifungal and antitubercular activity of (the roots of) Pelargonium reniforme (CURT) and Pelargonium sidoides(DC) (Geraniaceae) root extracts. S Afr J Bot 2006;72: 232-7.

Moyo M, Van Staden J. Medicinal properties and conservation of Pelargonium sidoides DC. J Ethnopharmacol 2014;152: 243– 55.