ADRİAMİSİNİN OLUŞTURDUĞU TESTİS HASARI ÜZERİNE E VİTAMİNİ VE SELENYUM’UN KORUYUCU ETKİSİNİN İNCELENMESİ

Year 2021, , 1 - 8, 09.04.2021

Mehmet Alparslan Ünal , Esra Balcıoğlu , Pınar Bilgici , Saim Özdamar

https://doi.org/10.34108/eujhs.781510

Abstract

Adriamisin, kanser tedavisinde yaygın olarak kullanılan bir kemoterapi ilacıdır. Adriamisin kanser hücreleri yanında testis dahil birçok dokuda zararlı etki göstermektedir. Selenyum ve Vitamin E üreme organları ve kısırlık üzerine koruyucu özelliklere sahiptirler. Bu çalışmanın amacı, Adriamisin ile indüklenen testis hasarına karşı E vitamini ve Selenyumun koruyucu etkisini araştırmaktır. Çalışmada 64 erkek sıçan her grupta 8 adet olacak şekilde; Kontrol, Adriamisin, Vitamin E, Selenyum, Vitamin E+Selenyum, Adriamisin+Vitamin E, Adriamisin+Selenyum, Adriamisin+VitaminE+Selenyum gruplarına ayrıldı. Testis dokuları eksize edildi ve %10’lukformaldehit içinde fikse edilerek rutin histolojik doku takibi basamaklarından geçirilerek parafin bloklara gömüldü. 5µm kalınlığındaki kesitler alınarak histopatolojik değerlendirme için Hematoksilen&Eozin ile boyanarak ışık mikroskobunda incelendi. Seminifer tübüllerdeki hasar JTBS ile belirlendi. Biyokimyasal yöntemlerle, Malondealdehit, Süperoksit Dismutaz, Katalaz ve Glutatyon Peroksidaz aktivitelerine bakıldı. Daha sonra kandaki serum örneklerinden testesteron seviyeleri ölçümlendi. TUNEL yöntemiyle apoptotik hücrelerin sayımı yapıldı. Sonuç olarak, dokular histopatolojik olarak değerlendirildiğinde diğer gruplardan farklı olarak ADR grubuna ait testis dokusunda düzensizlikler gözlendi. TUNEL yöntemi sonuçlarına göre ADR grubunda TUNEL pozitif hücre sayısı diğer gruplarla kıyaslandığında artış tespit edilmiştir (p=0.001). Testis dokusundaki MDA (p=0.593), CAT (p=0.469), GPx (p=0.655) ve SOD (p=0.907) değerleri kıyaslandığında ise gruplar arasında anlamlı bir farklılık bulunmamıştır. Testosteron düzeyleri, ADR uygulanan tüm gruplarda daha düşük bulunmuştur. Bu bağlamda gruplar arasında testosteron düzeyleri açısından anlamlı fark ortaya çıkmıştır (p=0.001). Adriamisin uygulanması sıçan testis dokularında hasar oluşturmuştur. Vitamin E ve Selenyum tedavisinin testis dokularında kısmen düzeltici etkiye sahip olduğunu kanıtlamıştır.

Keywords

Adriamisin, Selenyum, Sıçan, Testis, Vitamin E

Supporting Institution

Erciyes Üniversitesi Bilimsel Araştırmalar Projesi Birimi

Project Number

TYL-2016-6937

INVESTIGATION OF PROTECTIVE EFFECTS OF SELENIUM AND VITAMIN E ON ADRIAMYCIN-INDUCED TESTICULAR INJURY

Abstract

Adriamycin is a widely used chemotherapy drug in the treatment of cancer. Adriamycin has harmful effects on many tissues including testis as well as cancercells. Selenium and Vitamin E have reproductive organs and protective properties on infertility. The aim of this study to investigate the protective effect of Vitamin E and Selenium against Adriamycinin duced testicular injury. In this study, 64 male rats were 8 in each group; Control, Adriamycin, Vitamin E, Selenium, Vitamin E+Selenium, Adriamycin+Vitamin E, Adriamycin+Selenium, Adriamycin+VitaminE+Selenium divided in to groups. The testis tissues were excised and fixed in 10% formaldehyde and embedded in parafin blocks through routine histological follow-up steps. Sections of 5 µm thickness were stained with Hematoxylen&Eosin for histopathological evaluation and studied underlight microscope. The damage in the seminiferous tubules was determined through JTBS. Malondealdehyde, Superoxide Dismutase, Catalase and Glutathione Peroxidase activities were investigated by biochemical methods. Testosterone levels were measured from the serum samples. Apoptotic cells were counted by the TUNEL method. As a result, when the histopathological evaluation of the tissues was completed, irregularities were observed in the testicular tissues of the ADR group, unlike the other groups. According to the results of the TUNEL method, the number of TUNEL positive cells in the ADR group was increased compared to the other groups (p=0.001). When the values of MDA (p = 0.593), CAT (p = 0.469), GPX (p = 0.655) and SOD (p = 0.907) were compared in testicular tissue, nosignificant difference was found between the groups. However, testosterone levels were lower in all ADR groups compared to the other groups and this decrease was statistically significant. Testosterone levels were found to be lower in all ADR groups. In this context, there was a significant difference between the groups in terms of testosterone levels (p = 0.001). As a result, Adriamycin administration caused damage to rat testicular tissue. Vitamin E and Selenium treatment have proven to have a partial corrective effect on testicular tissues.

Keywords

Adriamycin, Rat, Selenıum, Testes, Vitamin E

Project Number

TYL-2016-6937

References

• 1. Pichiah PB, Sankarganesh A, Kalaiselvi S, Indirani K, Kamalakkannan S, SankarGanesh D, et. al Adriamycin induced spermatogenesis defect is due to the reduction in epididymal adipose tissue mass: A possible hypothesis. Med Hypotheses 2012; 78(2): 218-20.
• 2. Potnuri AG, Kondru SK, Samundrala PK, Allakonda L. Prevention of adriamycin induced cardiotoxicity in rat – A comparative study with sub acute angiotensin-converting enzyme inhibitorand non selective beta blocker therapy. Met. &Endorcine 2017; 14: 59-64.
• 3. Lobo V, Patil A, Phatak A, Chandra N. Free radicals, antioxidants and functional foods: Impact on human health. Pharmacogn Rev 2010; 4(8): 118-126.
• 4. Rengaraj D, Hong YH. Effects of dietary vitamin E on fertility functions in poultry species. Int.j. Mol. Sci 2015; 16(5): 9910-21.
• 5. Sakr SA, Mahran HA, Nofal AE. Effect of selenium on carbimazole-induced testicular damage and oxidative stress in albino rats. J Trace Elem Med Biol 2011; 25(1): 59-66.
• 6. Rayman MP. The importance of selenium to human healt. Lancet 2000; 356(9225): 233-41.
• 7. Naderi M, Keyvanshokooh S, Salati AP, Ghaedi A. Combined or individual effects of dietary vitamin E and selenium nanoparticles on humoral immune status and serum parameters of rainbow trout (Oncorhynchus mykiss) under high stocking density. Aquaculture 2017; 474: 40-47.
• 8. Bernabucci U, Ronchi B, Lacetera N, Nardone A. Markers of oxidative status inplasma and erythrocytes of transition dairy cows during hot season. J Dairy Sci 2002; 85(9): 2173–9.
• 9. Hernken RW, Harmon RJ, Tramsmel S. Selenium for Dairy Cattle: a Role for Organic Selenium. In: Lyons TP, Jacques KA, editors. Biotechnology in feed industry. Proceedings of the Alltech 14th annual symposium. Loughborough, LEC, UK: Nottingham University Press; 1998; 797-803.
• 10. El-Shenawy NS, AL-Harbi MS, Hamza RZ. Effect of vitamin E and selenium separately and in combination on biochemical, immunological and histological changes induced by sodium azide in male mice. Exp Toxicol Pathol 2015; 67(1): 65-76.
• 11. Chemotherapy: Maging side effects and safe handling. Can Vet J 2009; 50: 665-668.
• 12. Mut-Salud N, Álvarez PJ , Garrido JM , Carrasco E, Aránega A , Rodríguez-Serrano F. Antioksidant intake and antitumor therapy: Toward nutritional recommendations for optimal results. Oxid Med Cell Longev 2016; 2016: 2-56.
• 13. Singh KC, Kaur R, Marar T. Ameliorative Effect of Vitamin E on Chemotherapy Induced Side Effects in Rat Liver. Jpt 2011; 5(6): 481-492.
• 14. Boussada M, Ali RB, Said AB, Bokric K, Akachac AB, Dzirib C, et al. Selenium and a newly synthesized thiocyanoacetamide reduce doxorubicin gonadotoxicity in male rat. Biomed.& Pharmacotherapy 2017; 89: 1005-1017.
• 15. Blum RH, Carter SK. Adriamycin. A new anticancer drug with significant clinical activity. Ann İntern Med 1974; 80(2): 249-59.
• 16. Herman EH, Ferrans VJ. Animal models of anthracycline cardiotoxicity: basic mechanisms and cardioprotective activity. Prog Pediatr Cardiol 1997; 8(2): 49–58.
• 17. Au WW, Hsu TC. The genotoxic effects of adriamycin in somatic and germinal cells of the mouse. Mutat Res 1980; 79(4): 351–61.
• 18. Lui RC, Laregina MC, Herbold DR, Johnson FE. Testicular cytotoxicity of intravenous doxorubicin in rats. J Urol 1986; 136(4): 940–43.
• 19. Kato M, Makino S, Kimura H, Ota T, Furuhashı T, Nagamura Y. Sperm motion analysis in rats treated with adriamycin and its applicability to male reproductive toxicity studies. J Toxicol Sci 2001; 26(1): 51-9.
• 20. Çeribaşı AO, Sakin F, Türk G, Sönmez M, Ateşşahin A. Impact of ellagic acid on adriamycin-induced testicular histopathological lesions, apoptosis, lipid peroxidation and sperm damages. Exp Toxicol Pathol 2012; 64(7-8): 717-24.
• 21. Türk G, Çeribaşı AO, Sakin F, Sönmez M, Ateşşahin A. Antiperoxidative and antiapoptotic effects of lycopene and elagic acid on cyclophosphamide-induced testicular lipid peroxidation and apoptosis. Reprod Fertil Dev 2010a; 22(4): 587-96.
• 22. Mohamed RH, Karam RA, Hagrass HA, Amer MG, Abd El-Haleem MR. Antiapoptotic effect of spermatogonial stem cells on doxorubicin-induced testicular toxicity in rats. Gene 2015; 561(1): 107-14.
• 23. Rizk SM, Zaki HF, Mina MA. Propolis Attenuates Doxorubicin- Induced Testicular Toxicity in Rats. Food Chem. Toxicol 2014; 67: 176-86.
• 24. Niki E. Evidence for beneficial effects of vitamin E. Korean J. Intern. Med 2015; 30(5): 571–79.
• 25. EFSA Panel on Dietetic Products NaAN: Scientific opinion on the substantiation of health claims related to vitamin E and protection of DNA, proteins and lipids from oxidative damage (ID 160, 162, 1947), maintenance of the normal function of the immune system (ID 161, 163). EFSA J 2010; 8: 1816–1846.
• 26. Gurel A, Coskun O, Armutcu F, Kanter M, Ozen OA. Vitamin E against oxidative damage caused by formaldehyde in frontal cortex and hippocampus: biochemical and histological studies. J Chem Neuroanat 2005; 29(3): 173–8.
• 27. El-Faras AA, Sadek IA, Ali YE, Khalil M, Mussa EB. Protective effects of Vitamin E on CCl4- induced testicular toxicitiy in mle rats. Physiol Int 2016; 103(2): 157-168.
• 28. Momeni HR, Oryan S, Eskandari N. Effect of vitamin E on sperm number and testis histopathology of sodium arsenite-treated rats. Reprod Biol 2012; 12(2): 171-81.
• 29. Yin HP, Xu JP, Zhou XQ, Wang Y. Effects of vitamin E on reproductive hormones and testis structure in chronic dioxin-treated mice. Toxicol. Ind. Health 2012; 28(2): 152–161.
• 30. Song R, Yao X, Shi L, Ren Y, Zhao H. Effects of dietary selenium on apoptosis of germ cells in the testis during spermatogenesis in roosters. Theriogenology 2015; 84(4): 583-8.
• 31. Shi LG, Yang RJ, Yue WB, Xun WJ, Zhang CX, Ren YS et. al. Effect of elemental nano-selenium on semen quality, glutathione peroxidase activity, and testis ultrastructure in male Boer goats. Anim Reprod Sci 2010; 118(2-4): 248-54.
• 32. Olson GE, Winfrey VP, Hill KE, Burk RF. Sequential development of flagellar defects in spermatids and epididymal spermatozoa of selenium-deficient rats. Reproduction 2004; 127(3): 335-42.
• 33. Li JL, Gao R, Li S, Tang ZX, Xu SW. Testicular toxicity induced by dietary cadmium in cocks and ameliorative effect by selenium. Biometals 2010; 23(4): 695-705.
• 34. El-Maraghy SA, Nassar NN. Modulatory effects of lipoic acid and selenium against cadmium-induced biochemical alterations in testicular steroidogenesis. J. Biochem. Mol. Toxicol 2011; 25(1): 15-25.
• 35. Seema P, Swathy SS, Indira M. Protective effect of Selenium on Nicotine-Induced Testicular Toxicitiy in Rats. Biol Trace Elem Res 2007; 120(1-3): 212-8.
• 36. Cao Z, Shao B, Xu F, Liu Y, Li Y, Zhu Y. Protective effect of Selenium on Aflotoxin B1-Induced Testicular Toxicitiy in Mice. Biol Trace Elem Res 2017; 180(2): 233-238.
• 37. Kara H, Cevik A, Konar V, Dayangac A, Yilmaz M. Protective effects of antioxidants against cadmium-induced oxidative damage in rat testes. Biol Trace Elem Res 2007; 120(1-3): 205-11.
• 38. Taghizadeh L, Eidi A, Mortazavi P, Rohani AH. Effect of selenium on testicular damage induced by varicocele in adult male Wistar rats. J Trace Elem Med Biol 2017; 44: 177-185.
• 39. Gan L, Liu Q, Xu HB, Zhu YS, Yang XL. Effects of selenium overexposure on glutathione peroxidase and thioredoxin reductase gene expressions and activities. Biol Trace Elem Res 2002; 89(2): 165-75.
• 40. Amara IB, Soudani N, Hakim A, Troudi A, Zeghal KM, , Boudawara T, et al. Selenium and vitamin E, natural antioxidants, protect rat cerebral cortex against dimethoate-induced neurotoxicity. Pesticide Biochemistry and Physiology 2011; 101(3): 165-174.