Cu/Zn superoxide dismutase enzyme immunoreactivity in the stomach tissue of rats fed with mussels (Mytilus galloprovincialis)

Abstract

Objectives: Mussels accumulate heavy metals in their tissues. Although there are few data about the toxicity of seafood that is exposed to environmental pollution, there are no animal studies about the gastric toxicity of mussels grown in the Dardanelles. The antioxidant Cu/Zn superoxide dismutase (Cu/Zn SOD) enzyme catalyzes the hydrogen peroxide dismutation of superoxide radicals and removes the effects of free radicals which cause oxidative stress. The purpose of the study was to demonstrate the Cu/Zn SOD in the stomach tissues of rats which are fed with mussels that are collected from the Çamburnu region of the Dardanelles. 

Methods: A total of 24 male Wistar albino rats were randomly divided into four groups: Group 1 (n = 6), control group fed with standard rat food; Group 2 (n = 6), 75% mussels and 25% standard rat food daily; Group 3 (n = 6), 75% mussels and 25% standard rat food every two days; and Group 4 (n = 6), 75% mussels and 25% standard rat food every three days. To detect Cu/Zn SOD localization in the tissues, the LAB-SA Detection System was used. 

Results: Cu/Zn SOD enzyme immunoreactivity was not detected in Group 1 and in samples without Cu/Zn SOD primer antibody. Cu/Zn SOD enzyme immunoreactivity was detected 82% in Group 2, 79% in Group 3, and 61% in Group 4. There was statistically significant difference between the Cu/Zn SOD immunoreactivity of epithelial cells in the gastric mucosa of the rats in the experimental and control groups (p < 0.05). 

Conclusions: Determination of the increase of Cu/Zn SOD enzyme in the gastric mucosa of mussel-fed rats that we used in our study suggests that it triggers the antioxidant defense mechanism against mussel toxicity.

Keywords

• Immunohistochemistry,Dardanelles,mussel,Cu/Zn superoxide dismutase,stomach

References

1. [1] Julshamn K, Grahl-Nielsen O. Distribution of trace elemets from industrials discharges in the Hardangerfjord, Norway. A multivariate data-analysis of saithe, flounder, and blue mussel as sentinel organisms. Mar Pollut Bull 1996;32;564-71.
2. [2] Bat L, Gündoğdu A, Öztürk M, Öztürk M. Copper, zinc, lead and cadmium concentrations in the Mediterranean mussel Mytilus galloprovincialis Lamarck, 1819 from the Sinop coast of the Black Sea. Turk J Zool 1999;23:321-6.
3. [3] Gezen MR, Demir N, Çetin M, Ballı M. Çanakkale Boğazı’ndaki deniz kestanesi (Paracentrotuslividus)’nin ağır metal düzeyleri üzerinde bir araştırma. Ekoloji Sempozyumu, P2:10, Düzce, 2011.
4. [4] IARC (International Agency for Research on Cancer). Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to Humans: Arsenic and Arsenic Compounds (Group 1), vol. 7, World Health Organization, Lyon, France, 1987;100-6.
5. [5] Ibrahim D, Froberg B, Wolf A, Rusyniak DE. Heavy metal poisoning: clinical presentations and pathophysiology. Clin Lab Med 2006;26:67-97.
6. [6] Valko M, Morris H, Cronin MT. Metals, toxicity and oxidative stress. Curr Med Chem 2005;12:1161-208.
7. [7] Mates JM, Perez-Gomez C, Nunez de Castro I. Antioxidant enzymes and human diseases. Clin Biochem 1999;32:595-603.
8. [8] Fridovich I. Superoxide radical and superoxide dis mutases. Annu Rew Biochem 1995;64:97-112.

9. [9] Sandolio LM, Lopez-Huertas E, Bueno P, Del Rio LA. Immunocytochemical localization of copper, zinc superoxide dismutase in peroxisomes from water-melon (Citrullus vulgaris Schrad) cotyledons. Free Radic Res 1997;26:187-94.
10. [10] Teixeria HD, Schumacher RI, Meneghini R. Lower intracellular hydrogen peroxide levels in cells over-expressing CuZn-superoxide dismutase. Proc Natl Acad Sci U S A 1998;95:7872-5.
11. [11] Chitra KP, Pillai KS. Antioxidants in health. Ind J Physiol Pharmacol 2002;46:1-5.
12. [12] Worthington Enzyme Manual. Worthington Biochemical Corporation. Retrieved 2009;03-01.
13. [13] Tosun E, Tosun M, Avunduk MC. [Usage of cell proliferatıon markers as a prognostic factor in breast infiltrative ductal carcinoma]. Optimal Tıp Dergisi 2006;19:27-40. [Article in Turkish]
14. [14] Bakır K, Kapucuoğlu N, Bülbül HD. PCNA (Proliferating Cell NuclerarAntigene) immunohistokimyasal boyanması ve AgNORs’un meme karsinomunda prognozu belirlemedeki rolü. Türk Patholoji Dergisi 1996;12:35-8.
15. [15] Avunduk MC, Tavlı Ş, Yol S, Tavlı L, Yavuz A, Güngör S, et al. [Using of PCNA, Ki-67 and AgNOR as a markers of cell proliferation in adenocarcinomas of stomach]. Ankara Üniversitesi Tıp Fakültesi Mecmuası 2000;53:11-5. [Article in Turkish]
16. [16] Gezen MR, Demir N, Çetin M. Çanakkale Boğazı’nın Umurbey kıyısındaki deniz suyu ve bazı yumuşakçalarda (Bivalvia ve Gastropoda) ağır metal düzeylerinin araştırılması. Ekoloji Sempozyumu, P2: 11, Düzce, 2011.
17. [17] Demir N, Gezen MR, Ballı M. Çanakkale Boğazı’nın Karacaören kıyısındaki deniz suyu ve bazı yumuşakçalarda (Bivalvia ve Gastropoda) ağır metal düzeylerinin araştırılması. Ekoloji Sempozyumu, P1:39, Düzce, 2011.
18. [18] Rainbow PS. Ecophysiology of trace metal uptake in crustaceans. Estuar Coast Shelf Sci 1997;44:169-75.
19. [19] Paydaş S. [Heavy metal nephrotoxicity]. Turkiye Klinikleri J Nephrol-Special Topics 2016;9:60-8. [Article in Turkish]
20. [20] Krishnamurthy P, Wadhwani A. Antioxidant enzymes and human health. In: Antioxydant Enzyme. Amr El-Missiry M, ed. chapter 1. IntechOpen Ltd. London, 2012;pp.1-15.
21. [21] Vang O, Rasmussen BF, Andersen O. Combined effects of complex mixture of potentially anti-carcinogenic compounds on antioxidant anzymes and carcinogen metabolizing enzymes in the rat. Cancer Lett 1997;114:283-6.
22. [22] Gezen MR. Histopathological research and diagnosis of important paiting methods. JSP 2017;1:23-7.