Ergul Belge KURUTAŞ, Figen DORAN, Yakup GÜMÜŞALAN, Yalçın ATLI

Sağlıklı görünen farelerin karaciğer ve meme dokularında oksidatif stres biyobelirteçlerinin düzeyleri

Giriş: Bu çalışmada ileride yapılacak toksikolojik araştırmalara temel veri sağlamak amacıyla karaciğer ve meme dokularında antioksidan sistemler [glukoz-6-fosfat dehidrogenaz (G6PDH), katalaz (CAT), süperoksit dismutaz (SOD), glutatyon-S-transferaz (GST), redükte glutatyon (GSH)] incelendi. Buna ek olarak, lipit peroksidasyonun bir göstergesi olarak karaciğer ve meme dokularında tiyobarbiturik asit reaktif maddesi (TBARS) ölçüldü. Materyal ve Metod: Sağlıklı görünen, histopatolojik inceleme sonucu normal karaciğer ve meme dokusu bulgusu bulunan 66 fare (36 erkek, 30 dişi) çalışmaya alındı. Dokular soğuk % 1,15 KCI ile homojenize edildi. Antioksidan enzimler, GSH ve TBARS düzeyi 14.000 rpm"de santrifüjden sonra elde edilen süpernatant"ta spektrofotometrik olarak ölçüldü. Bulgular: Erkek ve dişi farelerin karaciğer dokularındaki antioksidan sistemlerin ve TBARS düzeylerinin meme dokusundan yüksek olduğu gözlendi (P < 0,05). Bununla birlikte, meme dokusundaki G6PDH hariç CAT, SOD, GST, GSH ve TBARS düzeylerinin erkek ve dişi fareler arasında farklılık göstermediği saptandı (P > 0,05). Öte yandan, erkek fare karaciğerinde G6PDH hariç CAT, SOD, GST, GSH ve TBARS düzeylerinin dişi karaciğerine göre daha yüksek bulundu (P < 0,05). Sonuç: Sonuçlar, karaciğer dokusunda antioksidan savunmanın meme dokusuna oranla yüksek olmasının, karaciğerde olası çeşitli toksik maddelere karşı kompensatuar yanıta bağlı olabileceğini ve bu şekilde hücreleri oksidatif hasara karşı koruyabileceğini göstermiştir.

Anahtar Kelimeler:

Antioksidan sistemler, TBARS, karaciğer, meme

The levels of oxidative stress biomarkers of liver and mammary tissues of apparently mice.

Objective: In the present study, the levels of antioxidant systems such as glucose-6-phosphate dehydrogenase (G6PDH), catalase (CAT), superoxide dismutase (SOD), glutathione-S-transferase (GST) and, also reduced glutathione (GSH) in liver and breast tissues were examined in order to obtain basal data for subsequent toxicological investigations. Additionally, the level of thiobarbituric acid reactive substance (TBARS) was measured in liver and breast tissues as an index of lipid peroxidation. Materials and Methods: Sixty-six apparently healthy mice (36 male, 30 female) had normal liver and breast tissues (histopathological data) were taken into the study. The tissues were homogenised with ice-cold 1.15 % KCI. The activities of antioxidant enzymes, the levels of GSH and TBARS were measured as spectrophotometric in the supernatant obtained from centrifugation at 14.000 rpm. Results: The levels of antioxidant systems and TBARS in liver were significantly higher than those found in breast tissue of female and male mice (P < 0.05). However, the levels of CAT, SOD, GST, GSH and TBARS in breast tissue except G6PDH enzyme were not significantly different between female and male mice (P > 0.05). On the other hand, the levels of CAT, SOD, GST, GSH and TBARS in male liver except G6PDH enzyme were higher than female liver (P < 0.05). Conclusion: Results suggest that antioxidant defense of the liver tissue was higher compared to breast tissue possibly due to a compensatory response to various toxic substances in liver and thereby protects the cells against oxidative damage.

Keywords:

Antioxidant systems, TBARS, liver, breast,

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Ergün Y, Kurutaş EB, Ozdil B, Güneşaçar R. Evaluation of nitrite/nitrate levels in relation to oxidative stress parameters in liver cirrhosis. Clin Res Hepatol Gastroenterol. 2011; 35: 303-8.

Prohaska JR, Sunde RA. Comparison of liver glutathione peroxidase activity and mRNA in female and male mice and rats. Comp Biochem Physiol. 1993; 105:111-6.

Harris ED. Regulation of antioxidant enzymes. FASEB J. 1992; 6: 2675-83.

Martini G, Toniolo D, Vulliamy T et al. Structural analysis of the x-linked gene encoding human glucose 6- phosphate dehydrogenase. EMBO J. 1986; 5: 1849-55.

Georgeson GD, Szony BJ, Streitman K, Varga IS, Kovács A, Kovács L, László A. Antioxidant enzyme activities are decreased in preterm infants and in neonates born via caesarean section. Eur J Obstet Gynecol Rep Biol. 2002; 103:136-9.

Akman SA, Forrest G, Chu FF, Doroshow JH. Resistance to hydroperoxide associated with altered catalase mRNA stability in MCF7 breast cancer cells. Biochim Biophys Acta. 1989; 1009: 70-4.

Singhal SS, Saxena M, Ahmad H, Awasthi YC. Glutathione S-transferase of mouse liver: sexrelated differences in the expression of various isozymes. Biochim Biophys Acta. 1992; 1116: 137

Hussey AJ, Hayes JD. Human Mu-class glutathione S-transferases present in liver,skeletal muscle and testicular tissue. Biochim Biophys Acta. 1993; 1203: 131Jakoby WB, Habig WH, Keen JH et al. Glutathione S- transferases: Catalytic aspects. In: Arias IM, Jacoby WB, eds. Glutathione Metabolism and Function. Raven Press. New York. 1976; 189-211.

Stohs SJ, Al-Turk WA, Angle CR, Heinicke RJ. Glutathione-S-transferase activity in liver, lung and intestinal mucose of aging female mice. Gen Pharmacol. 1982; 13: 519-22.

Stohs SJ, Hassing JM, Al-Turk WA. Glutathione levels in hepatic and extrahepatic tissues of mice as a function of age. Age. 1980; 3: 11-5.

Nakai A, Oya A, Kobe H, Asakura H, Yokota A, Koshino T, Araki T. Changes in maternal lipid peroxidation levels and antioxidant enzymatic activities before and after delivery. J Nippon Med Sch. 2000; 67: 434-9.

Fridovich I. Superoxide radical: an endogenous toxicant. Ann Rev Pharmacol Toxicol. 1983; 23: 239

Beutler E. Red Cell Metabolism: A Manual of Biochemical Methods, 3rd ed. Grune & Stratton. New York. 1984; 105-6.

Mannervik B, Guthenberg C. Glutathione-Stransferase (human placenta). Meth Enzymol. 1981; 77: 231-5.

Ohkawa H, Ohishi N, Tagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 1979;95: 351-8.

Lowry OH, Rosenbrough NJ, Farr ALL, Randall RJ. Protein measurement with the folin phenol reagent. J Biol Chem 1951;193: 265-75.

Bancroft JD, Stevens A. Theory and Practice of Histological Techniques, 5th ed. Edinburg: Churchill Livingstone; 2001; 1-15.

Lawrence MS, Jefferson L, James T, et al. Some comparative aspects of the organ distribution of superoxide dismutase activity in the freshwater turtle, Psedemysscripta elegans. Comp Biochem Physiol. 1977; 58:377-9.

Winzer K, Van Noorden CJ, Köhler A. Glucose-6phosphate dehydrogenase: the key to sex-related xenobiotic toxicity in hepatocytes of European flounder. Aquat Toxicol. 2002; 56: 275-88,.

Martins RN, Hartmann PE, Stokes GB. Comparative profiles of the hexose monophoshate dehydrogenases in rat tissues over the lactation cycle. Aust J Biol Sci. 1985;38: 295-303.

Rebrin I, Kamzalov S, Sohal RS. Effects of age and caloric restriction on glutathione redox state in mice. Free Radic Biol Med. 2003; 35: 626-35.

Chang HL, Holten D, Karin R. Distribution of the multiple molecular forms of glucose-6-phosphate dehydrogenase in different physiological states. Can J Biochem. 1979; 57:396-401.

Grigor MR, Geursen A, Sneyd MJ, Warren SM. Regulation of lipogenic capacity in lactating rats. Biochem J. 1982;.208: 611-8.

Schisler NJ, Singh SM. Inheritance and expression of tissue-specific catalase activity during development and aging in mice. Genome 1987;29:748-60.

Takenaka T, Goto F. Alteration of lipid peroxidation and the activity of peroxide metabolism enzymes in the liver, kidney and lung following the administration of araquat in mice. Masui. 1994; 43: 34-40.

Nakao C, Ookawara T, Sato Y, Kizaki T, Imazeki N, Matsubara O, et al.Extra-cellular superoxide dismutase in tissues from obese (ob/ob) mice. Free Radic Res. 2000; 33:229-41.

Izokun-Etiobhio BO, Oraedu ACI, Ugochukwu EN. A comparative study of superoxide dismutase in various animal species. Comp Biochem Physiol. 1990; 95: 521-3.

Takenouchi Y, Kobayashi T, Matsumoto T, Kamata K. Gender differences in age-related endothelial function in the murine aorta. Atherosclerosis. 2009; 206:397-404.

Draper HH, Squires EJ, Mahmoodi H, Wu J, Agarwal S, Hadley M. A comparative evaluation of thiobarbituric acid methods for the determination malondialdehyde in biological materials. Free Radic Biol Med 1993;15: 353-63.

Sharma R, Ahmad H, Singhal SS, Saxena M, Srivastava SK, Awasthi YC. Comparative studies on the effect of butylated hydroxyanisole on glutathione and glutathione S-transferases in the tissues of male and female CD-1 mice. Comp Biochem Physiol. 1993; 105: 31-7.

Hovey RC, Trott JF, Vonderhaar BK. Establishing a framework for the functional mammary gland: from endocrinology to morphology. J Mammary Gland Biol Neoplasia. 2002; 7:17-39.

Yazışma Adresi / Address for Correspondence: Dr.Ergül Belge Kurutaş Kahramanmaraş Sütçü İmam Üniversitesi Biyokimya Anabilim Dalı Yörük Selim mah., Hastane cad., No: 32 46050/ KAHRAMANMARAŞ Tel: 0344 2212337/358 Faks: 0344 2212371 geliş tarihi/received :26.07.2012 kabul tarihi/accepted:21.08.2012