FİKRİYE POLAT, GÜNSEL BİNGÖL, NESRİN TURAÇLAR

An investigation of micronucleus induction by Butylated Hydroxytoluene in wistar rat bone marrow cells

Bu çalışma ile sentetik antioksidan ve yiyeceklerde katkı maddesi olarak kullanılan Bütil Hidroksitoluenin (BHT) rat kemik iliğinde mikronükleuslu polikromatik eritrositlerin sayılarında herhangi bir artışa neden olup olmadığı araştırıldı. Mısır yağında çözülen BHT, iki farklı zaman periyodu ve üç farklı dozda (125, 250, 500 mg/kg v.a.), 8-10 haftalık erkek ve dişi Wistar albino sıçanlara (n=36) intraperitonal olarak verildi. BHT muamelesinden 12 ve 24 saat sonra kemik iliği örnekleri MNPCE sayısı için analiz edildi. Ayrıca, polikromatik eritrositlerin normokromatik eritrositlere oranı (PCE/NCE) değerlendirilerek BHT’nin kemik iliğindeki sitotoksik etkisine bakıldı. Bütün BHT dozları ve iki farklı zaman periyodunda MNPCE’lerin sayısında 1.9- 2.84 kat önemli artışlar bulundu. Diğer taraftan BHT, kemik iliği sitotoksisite belirteci olan PCE/NCE oranını kontrol grubuna kıyasla düşürdü. Sonuç olarak bu çalışmada BHT’nin rat kemik iliğinde MNPCE oluşumunu artırdığını ve bu artışın, uygulanan dozlar ve verilme şekliyle ilişkili olabileceğini düşünmekteyiz.

Bütil Hidroksitoluen tarafından mikronükleus indüksiyonunun wistar sıçan kemik iliği hücrelerinde araştırılması

This study aims to investigate whether Butylated Hydroxytoluene (BHT), the synthetic antioxidant and food additive, increases the frequency of micronucleated polychromatic erythrocytes (MNPCEs) in rat bone marrow. BHT, dissolved in corn oil, was administered intraperitoneally to 8-10 week old male and female Wistar rats (n=36) in three different doses (125, 250 and 500 mg/kg b.w.) for two different time periods. 12- and 24-h after BHT treatment, the bone marrow samples were analyzed for the frequency of MNPCEs. Additionally, by evaluating the ratio of polychromatic erythrocyte to normochromatic erythrocyte (PCE/NCE), the cytotoxic effect of BHT on bone marrow was tested. It was found that all BHT doses at two time periods significantly increased the MNPCEs frequency about 1.9-2.84 fold. On the other hand, BHT caused significant decreased in the PCE/NCE ratio, which is indicative of bone marrow cytotoxicity when compared to the control groups. This study showed that BHT increased the formation of MNPCEs in rat bone marrow and we think that this increase might be related to the applied doses and the administration way BHT.

PDF

___

1. Williams GM, Iatropoulos MJ, Whysner J: Safety assessment of butylated hydroxyanisole and butylated hydroxytoluene as antioxidant food additives. Food Chem Toxicol, 37, 1027-1038, 1999.
2. Polat F, Ozdemir O, Elagoz S: Analysis of Ki-ras exon 2 gene mutations In 3-methylcholanthrene and butylated hydroxytoluene-induced rat lung tissues. Turk J Biol, 32, 277-282, 2008.
3. Williams GM, Wang CX, Iatropoulos MJ: Toxicity studies of butylated hydroxyanisole and butylated hydroxytoluene. I. Genetic and cellular effects. Food Chem Toxicol, 28, 799-806, 1990.
4. Yoshida Y: Study on mutagenicity and antimutagenicity of BHT and its derivatives in a bacterial assay. Mutat Res, 242, 209-217, 1990.
5. Detringer SD, Torous DK, Tometsko AM: In vitro system for detecting non-genotoxic carcinogens. Environ Mol Mutagen, 21, 332-338, 1993.
6. Bomhard EM, Bremmer JN, Herbold BA: Review of the mutagenicity/ genotoxicity of butylated hydroxytoluene. Mutat Res, 277, 187-200, 2002.
7. Stanford Research Institute (S.R.I.): Report 14 submitted to U.S. Food and Drug Administration, 1972.
8. Jung KM, Ryu JC: Evaluation of the genetic toxicity of synthetic chemical (XVIII)-in vitro mouse lymphoma assay and in vivo supravital micronucleus assay with butylated hydroxytoluene (BHT). Mol Cell Toxicol, 23, 172-181, 2007.
9. World Health Organization, International Agency for Research on Cancer (WHO/IARC): Some Naturally Occurring and Synthetic Food Components, Furocoumarins and Ultraviolet Radiation, 40, 1998.
10. Schmid W: The micronucleus test. Mutat Res, 31, 9-15, 1975.
11. Agarwal DK, Chauhan LKS: An improved chemical substitute for fetal calf serum for the micronucleus test. Biotech Histochem, 68, 187-188, 1993.
12. Demirel C, Kilciksiz S, Ay OA, Gurgul S, Ay ME, Erdal N: Effect of N-acetylcysteine on radiation-induced genotoxicity and cytotoxicity in rat bone marrow. J Radiat Res, 50, 43-50, 2009.
13. Ledebur MV, Schmid W: The micronucleus test. Methodological aspects. Mutat Res, 19, 109-117, 1973.
14. Maier P, Schmid W: Ten model mutagens evaluated by the micronucleus test. Mutat Res, 40, 325-337, 1976.
15. Aksu P, Dogan A, Gul S, Kanici: Farelerde 3-Metilkolantren ile indüklenen Fibrosarkoma üzerine siteaminin etkileri: Genotoksisitenin araştırılması. Kafkas Univ Vet Fak Derg, 19 (6): 955-961, 2013. DOI: 10.9775/ kvfd.2013.9172
16. Celik A, Mazmanci B, Camlica Y, Askin A, Comelekoglu U: Induction of micronuclei by lambda-cyhalothrin in Wistar rat bone marrow and gut epithelial cells. Mutagenesis, 20, 125-129, 2005.
17. Sekeroglu V, Sekeroglu ZA: Micronucleus test for determining genotoxic damage. Türk Hijyen ve Deneysel Biyoloji Dergisi, 68, 241, 2011.
18. Olsen P, Meyer O, Bille N, Würtzen G: Carcinogenicity study on butylated hydroxytoluene in Wistar rats exposed in utero. Food Chem Toxicol, 24, 1-12, 1986.
19. EFSA Panel on Food Additives and Nutrient Sources Added to Food (ANS): Scientific Opinion on the re-evaluation of butylated hydroxytoluene BHT (E 321) as a food additive, EFSA J, 10, 2588, 2012.
20. Shelef LA, Chin B: Effect of phenolic antioxidants on the mutagenicity of aflatoxin B1, Appl Environ Microb, 40, 1039-1043, 1980.
21. Reddy BS, Hansen D, Mathews L: Effect of micronutrients, antioxidants and related compounds on the mutagenicity of 3,2’- dimethyl-4-aminobiphenyl, a colon and breast carcinogen. Food Chem Toxicol, 21, 129, 1983.
22. Budunova IV, Mittelman LA, Belitsky GA: Identification of tumor promoters by their inhibitory effect on intercellular transfer of Lucifer yellow. Cell Biol Toxicol, 5, 77-89, 1989.
23. Cicchetti R, Bari M, Argentin G: Induction of micronuclei in bone marrow by two pesticides and their differentiation with CREST staining: an in vivo study in mice. Mutat Res, 439, 239-248, 1999.
24. Lambert IB, Singer TM, Boucher SE, Douglas GR: Detailed review of transgenic rodent mutation assays. Mutat Res, 590, 1-280, 2005.
25. Yener Y, Dikmenli M: Increased micronucleus frequency in rat bone marrow after acrylamide treatment. Food Chem Toxicol, 47, 2120-2123, 2009.
26. Sekeroglu V, Sekeroglu ZA: Micronucleus test for determining genotoxic damage. Türk Hijyen ve Deneysel Biyoloji Dergisi, 68, 241, 2011.
27. Paschin YV, Bakhitova LM, Benthen TI: Increased antimutagenic activity of simple substituted phenols mixed with the hindered phenolic antioxidant dibunol. Food Chem Toxicol 24, 881-883, 1986.