Immunohistochemical distribution of glutathione peroxidase and its gene expression via RT-PCR in the liver tissue of melatonin-administered mice

The purpose of this study is to investigate the effect of exogenous administration of melatonin, which is a strong antioxidant agent, on the gene expression of glutathione peroxidase, which is one of the antioxidant enzymes, in the livers of mice and also to examine the immunohistochemical localization of the enzyme. The 24 male Swiss albino mice used in the study were divided into 3 groups: experimental (n = 8), sham (n = 8), and control (n = 8). A 10 mg/kg dose of melatonin was administered intraperitoneally to the experimental group for 4 weeks. Only ethanol and a serum physiological solution were applied to the sham group. No administration was done for the control group. It was observed that the expression level of the glutathione peroxidase 1 enzyme in the experimental group showed a statistically significant increase (P < 0.05) compared to the sham and control groups. In the immunohistochemical examinations, it was seen that immunoreactivity was especially intensified in the hepatocytes around Kiernan’s space and in the hepatocytes around the vena centralis, and that reaction was generally present both as cytoplasmic and nuclear reactions in the hepatocytes.

Anahtar Kelimeler:

Key words: Melatonin, liver, glutathione peroxidase 1, RT-PCR, immunohistochemistry

Immunohistochemical distribution of glutathione peroxidase and its gene expression via RT-PCR in the liver tissue of melatonin-administered mice

Keywords:

Key words: Melatonin, liver, glutathione peroxidase 1, RT-PCR, immunohistochemistry,

PDF

___

Nakane T, Asayama K, Kodera K, Hayashibe H, Uchida N, Nakazawa S. Effect of selenium deficiency on cellular and extracellular glutathione peroxidases: immunochemical detection and mRNA analysis in rat kidney and serum. Free Radic Biol Med 1998; 25: 504–511.
Akkuş İ. Serbest Radikaller ve Fizyopatolojik Etkileri. Konya, Turkey: Mimoza Yayınları; 1995 (book in Turkish).
Simmons TW, Jamall IS. Significance of alterations in hepatic antioxidant enzymes: primacy of glutathione peroxidase. Biochem J 1988; 251: 913–917.
Brzezinski A. Melatonin in humans. N Engl J Med 1997; 336: 186–195.
Yazıcı C, Köse K. Melatonin: Karanlığın antioksidan gücü. Erciyes Üniv Sağ Bil Derg 2004; 13: 56–65 (article in Turkish).
Ianăş O, Olinescu R, Bădescu I. Melatonin involvement in oxidative processes. Endocrinologie 1991; 29: 147–153.
Longoni B, Salgo MG, Pryor WA, Marchiafava PL. Effects of melatonin on lipid peroxidation induced by oxygen radicals. Life Sci 1998; 62: 853–859.
Kotler M, Rodriguez C, Sainz RM, Antolin I, MenendezPelaez A. Melatonin increases gene expression for antioxidant enzymes in rat brain cortex. J Pineal Res 1998; 24: 83–89.
Luna LG. Manual of Histologic Staining Methods of the Armed Forces Institute of Pathology. 3rd ed. New York, NY, USA: McGraw-Hill; 1968.
Hsu SM, Raine L, Fanger H. Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem 1981; 29: 577–580.
Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 1987; 162: 156–159.
Ferret PJ, Soum E, Negre O, Fradelizi D. Auto-protective redox buffering systems in stimulated macrophages. BMC Immunol 2002; 12: 3.
Kocamış H. Functional profiles of growth related genes during embryogenesis and postnatal development of chicken and mouse skeletal muscle. PhD, West Virginia University, Morgantown, WV, USA, 2001.
Yamamura M, Uyemura K, Deans RJ, Weinberg K, Rea TH, Bloom BR, Modlin RL. Defining protective responses to pathogens: cytokine profiles in leprosy lesions. Science 1991; 254: 277–279.
Minitab Inc. Minitab Reference Manual. Release 10 for Windows. State College, PA, USA: Minitab Inc.; 1994.
Tekin ME. Sağlık Bilimleri İçin Örneklerle Bilgisayarda Biyoistatistik. Konya, Turkey: Selçuk Üniversitesi Veteriner Fakültesi Yayın Ünitesi; 2003 (book in Turkish).
Banks WJ. Applied Veterinary Histology. 3rd ed. St. Louis, MO, USA: Mosby; 1993.
Deprem T. Sağlıklı ve diabet oluşturulmuş farelerin karaciğer dokusunda glutatyon peroksidaz enziminin immunohistokimyasal lokalizasyonu ve RT PCR ile gen ekspresyonu. PhD, Kafkas University, Kars, Turkey, 2009 (thesis in Turkish).
Asayama K, Yokota S, Dobashi K, Hayashibe H, Kawaoi A, Nakazawa S. Purification and immunoelectron microscopic localization of cellular glutathione peroxidase in rat hepatocytes: quantitative analysis by postembedding method. Histochemistry 1994; 102: 213–219.
Asayama K, Yokota S, Dobashi K, Kawada Y, Nakane T, Kawaoi A, Nakazawa S. Immunolocalization of cellular glutathione peroxidase in adult rat lungs and quantitative analysis after postembedding immunogold labeling. Histochem Cell Biol 1996; 105: 383–389.
Yoshimura S, Komatsu N, Watanabe K. Purification and immunohistochemical localization of rat liver glutathione peroxidase. Biochim Biophys Acta 1980; 621: 130–137.
Murakoshi M, Fukui N, Takekoshi S. Immunohistochemical and biochemical studies in 4-aminopyrazolopyrimidine (4-APP)-induced fatty liver. Tokai J Exp Clin Med 2002; 27: 73–
Asayama K, Dobashi K, Kawada Y, Nakane T, Kawaoi A, Nakazawa S. Immunohistochemical localization and quantitative analysis of cellular glutathione peroxidase in foetal and neonatal rat tissues: fluorescence microscopy image analysis. Histochem J 1996; 28: 63–71.
Rodriguez C, Mayo JC, Sainz RM, Antolín I, Herrera F, Martín V, Reiter RJ. Regulation of antioxidant enzymes: a significant role for melatonin. J Pineal Res 2004; 36: 1–9.
Lankoff A, Banasik A, Nowak M. Protective effect of melatonin against nodularin-induced oxidative stress. Arch Toxicol 2002; 76: 158–165.
Mayo JC, Sainz RM, Antoli I, Herrera F, Martin V, Rodriguez C. Melatonin regulation of antioxidant enzyme gene expression. Cell Mol Life Sci 2002; 59: 1706–1713.
Swiderska-Kołacz G, Klusek J, Kołataj A. The effect of melatonin on glutathione and glutathione transferase and glutathione peroxidase activities in the mouse liver and kidney in vivo. Neuro Endocrinol Lett 2006; 27: 365–368.
Bharti VK, Srivastava RS, Subramaian P, Warren Spence D, Pandi-Perumal SR, Brown GM. Cerebral epiphyseal proteins and melatonin modulate the hepatic and renal antioxidant defense of rats. Int J Nephrol 2011; 2011: 142896.
Mauriz JL, Molpeceres V, García-Mediavilla MV, González P, Barrio JP, González-Gallego J. Melatonin prevents oxidative stress and changes in antioxidant enzyme expression and activity in the liver of aging rats. J Pineal Res 2007; 42: 222–230.
Gómez M, Esparza JL, Nogués MR, Giralt M, Cabré M, Domingo JL. Pro-oxidant activity of aluminum in the rat hippocampus: gene expression of antioxidant enzymes after melatonin administration. Free Radic Biol Med 2005; 38: 104–111.