Protective Effects of Vitamin E against Methotrexate-Induced Testicular Damage in Rats: Histopathologic and flow cytometric study

Öz

Background: Methotrexate (MTX) can cause oxidative stress-related tissue damage.Vitamin E neutralizes lipid peroxidation arising from the effect of free oxygen radicals.In this study, the protective effect of vitamin E against possible MTX-related testicular damage was analyzed. Method: Thirty two mature male Spraque dawley rats were grouped as MTX, Vitamin E, MTX+Vitamin, Control groups. 20 mg/kg MTX intraperitoneal (i.p.) was applied to MTX Group in the first day; 100mg/kg i.p. vitamin E was applied to Vitamin E Group for 5 days; 20 mg/kg i.p. MTX in the first day and 100 mg/kg i.p. vitamin E for 5 days were applied to MTX+Vitamin E Group;2 ml physiological saline solution (i.p.) was applied to Control Group for 5 days. Histopathology, flow cytometry and apoptosis were evaluated on testicular tissue. Result: Apoptotic Index (%) and testicular damage were highest for MTX Group, and significant decrease was observed for MTX+Vitamin E Group compared to MTX Group.Seminiferous tubule size significant decreased in MTX Group and it increased in MTX+Vitamin E Group compared to MTX Group. No significant difference was found between MTX and MTX+Vitamin E Groups regarding germinal epithelium thickness and testicle weights. Conclusion: The results show that MTX can cause structural disruptions in testicles and vitamin E can rehabilitate MTX-related testicular damage.

Anahtar Kelimeler

• Methotrexate,Vitamin E,Apoptosis,TUNEL,Flow cytometry

Sıçanlarda Metotreksat ile İndüklenen Testis Hasarına Karşı E Vitamininin Koruyucu Etkileri: Histopatolojik ve Akım Sitometrik Çalışma

Öz

Amaç: Metotreksat oksidatif strese bağlı doku hasarına neden olmaktadır. E vitamini serbest oksijen radikallerinden etkisiyle oluşan lipt peroksidasyonunu nötralize eder. Bu çalışma ile E vitamininin olası Metotreksat bağlantılı testiküler hasara karşı koruyucu etkileri analiz edildi. Materyal-Metot: Otuz iki erkek Spraque dawley sıçan MTX, E Vitamini, MTX+ E Vitamini, Kontrol olarak gruplandırıldı. İlk gün MTX grubuna intraperitoneal(i.p.) 20 mg/kg MTX, E vitamini grubuna 5 gün 100 mg/kg i.p. E vitamini, MTX+E Vitamini grubuna ilk gün 20 mg/kg i.p. MTX ve 5 gün 100 mg/kg i.p. E vitamini uygulandı. Kontrol grubuna 5 gün 2 ml serum fizyolojik (i.p.) uygulandı. Testiküler dokuda histopatolojik, akım sitometrik analizler yapıldı ve apoptoz değerlendirildi. Bulgular: Apoptotik indeks (%) ve testiküler hasar en fazla MTX grubundayken, MTX+E Vitamini grubunda MTX grubuna göre anlamlı derecede azalma gözlendi. Seminifer tubule çapı MTX grubunda belirgin şekilde azalırken, MTX+E Vitamini grubunda MTX grubuna göre artış gözlendi. MTX ve MTX+E Vitamini grupları arasında germinal epitelyum ve testiküler ağırlıkta açısında anlamlı farklılık yoktu. Sonuç: Sonuçlara göre MTX testislerde yapısal hasara neden olabilirken, E vitamini MTX'e bağlı testiküler hasarı düzeltebilir.

Anahtar Kelimeler

• Metotreksat,E vitamin,Apopitozis,TUNEL,Akım sitometri

Kaynakça

1. 1. Panetta JC, Sparreboom A, Pui CH, Relling MV, Evans WE. Modeling mechanisms of in vivo variability in methotrexate accumulation and folate pathway inhibition in acute lymphoblastic leukemia cells. PLoS Comput Biol 2010; 2;6[12]:e1001019. doi: 10.1371/journal.pcbi.1001019.
2. 2. Chabner B, Wilson W, Supko J. Pharmacology and toxicity of antineoplastic drugs. Editorial: Lichtman MA, Beutler E, Seligsohn U, Kipps TJ, Kaushansky K. In Williams Hematology. 7th Edition. Mc Graw Hill Company, United States. 2007; 249-51.
3. 3. Van Ede AE, Laan RF, Blom HJ, De Abreu RA, Van De Putte LB. Methotrexate in rheumatoid arthritis: an update with focus on mechanisms involved in toxicity. Seminars in Arthritis and Rheumatism 1998; 27:277-292.
4. 4. Jollivet J., Cowan K., Curt G., Clendeninn N., Chabner B. The pharmacology and clinical use of methotrexate. N. Eng. J. Med, 1983;309:1094-1104.
5. 5. .Dutz JP, Ho VC. Immunosuppressive agents in dermatology. Dermatol Clin. 1998;16[2]:235-251.
6. 6. Kolli VK, Abraham P, Isaac B, Selvakumar D. Neutrophil infiltration and oxidative stress may play a critical role in methotrexate-induced renal damage. Chemotherapy. 2009; 55[2]:83-90.
7. 7. Miyazono Y, Gao F, Horie T. Oxidative stress contributes to methotrexate-induced small intestinal toxicity in rats. Scand J Gastroenterol, 2004; 39[11]:1119-27.
8. 8. Jahovic N, Çevik H, Şehirli AO, Yeğen BC, Şener G. Melatonin prevents methotrexate induced hepatorenal oxidative injury in rats. J Pineal Res. 2003; 34[4]: 282-287.

9. 9. Padmanabhan S, Tripathi DN, Vikram A, Ramarao P, Jena GB. Cytotoxic and genotoxic effects of methotrexate in germ cells of male Swiss mice. Mutat Res, 2008; 655[1-2]:59-67.
10. 10. Olson J. Klinik Farmakoloji. Birinci Baskı, Ankara, Hacettepe Taş Yayıncılık, 2000.
11. 11. Miketova P, Kaemingk K, Hockenberry M, Pasvogel A, Hutter J, Krull K, Moore IM. Oxidative changes in cerebral spinal fluid phosphatidylcholine during treatment for acute lymphoblastic leukemia. Biol Res Nurs, 2005; 6 [3]: 187-195.
12. 12. Brunton LL, Lazo JS, Parker KL [2006]. Goodman and Gilman’s the pharmacologycal basis of therapeutics, Mc Graw- Hill Companies, 2006, New-York.
13. 13. Bertram G. Katzung, Basic and Clinical Pharmacology, 9. Edition, Singapore: The McGraw-Hill Companies, 2004; 898-931.
14. 14. Işık A, Işılay L, Atabenli Erdemli E, Akbay C, Anafarta K. Sıçan Testisinde Metotreksat'ın Işık Ve Elektron Mikroskop Düzeyinde Etkileri. Ankara Üniversitesi Tıp Fakültesi Mecmuası 1997; Cilt 50, Sayı 3.
15. 15. Oktar S, Gokce A, Aydin M, Davarci M, Meydan S, Ozturk OH, Koc A. Beneficial effect of erdosteine on methotrexate-induced testicular toxicity in mice. Toxicology and Industrial Health. 2010; 26[7]: 433–438.
16. 16. Champe PC. Biyokimya. Çeviren: Gür E, Nobel Tıp Kitabevi, İstanbul, 1997; 438.
17. 17. Pekiner BD. Vitamin E as an antioxidant. J Fac Pharm Ankara, 2003; 32, 243-267.
18. 18. Leiber DC. The role of metabolizm in the antioxidant function of vitamin E. Crit Rev Toxicol, 1993; 23: 147-169.
19. 19. Nisbet H, Nisbet C, Akar A, Cevik M, Onder Karayigit M. Effects of exposure to electromagnetic field [1.8/0.9 GHz] on testicular function and structure in growing rats. Research in Veterinary Science. 2012; 93: 1001-1005.
20. 20. Silva AS, Villasenor RMV, Marquez SR, Gonzales MH, Jaime HB, Garcia XG, Montiel JLC. Testosterone levels and development of the penile spines and testicular tissue during the postnatal growth in Wistar Rats. Advances in Sexual Medicine, 2013; 3: 1-9.
21. 21. Shen H, Liao K, Wu HF, Lu HC, Li Y, Li Z, Zhang W. In utero exposure of high-dose di- n-butyl phthalate resulted in opposite effects on testicular cell apoptosis in late embryonic and pubertal male rat offspring. Hum Exp Toxicol, 2016. doi: 10.1177/0960327116685886.
22. 22. Rashed M, Ragab N, Shalaby A, Ragab W. Patterns Of Testicular Histopathology In Men With Primary Infertility. The Internet Journal of Urology.2007; 5: 2.
23. 23. Potten CS. What is an apoptotic index measuring? A commentary. Br J Cancer 1996; 74[11]:1743-8.
24. 24. Barbisan F, Motta Jde R, Trott A, Azzolin V, Dornelles EB, Marcon M, Algarve TD, Duarte MM, Mostardeiro CP, Unfer TC, Schott KL, da Cruz IB. Methotrexate-related response on human peripheral blood mononuclear cells may be modulated by the Ala16Val-SOD2 gene polymorphism. PLoS One; 2014; 9[10]:e107299. 25. Oufi HG, Al-Shawi NN. The effects of different doses of silibinin in combination with methotrexate on testicular tissue of mice. Eur J Pharmacol, 2014 ;5;730:36-40.
25. 26. Bangert CA, Costner MI. Methotrexate in dermatology. Dermatol Ther, 2007; 20[4]: 216-28.
26. 27. Sönmez MF, Çilenk KT, Karabulut D, Ünalmış S, Deligönül E, Öztürk İ, Kaymak E. Protective effects of propolis on methotrexate-induced testis injury in rat. Biomed Pharmacother. 2016; 79: 44-51.
27. 28. Sheikhbahaei F, Khazaei M, Rabzia A, Mansouri K, Ghanbari A. Protective Effects of Thymoquinone against Methotrexate-Induced Germ Cell Apoptosis in Male Mice. Int J Fertil Steril. 2016; 9[4]:541-7.
28. 29. Daggulli M, Dede O, Utangac MM, Bodakci MN, Hatipoglu NK, Penbegul N, Sancaktutar AA, Bozkurt Y, Türkçü G, Yüksel H. Protective effects of carvacrol against methotrexate-induced testicular toxicity in rats. Int J Clin Exp Med.2014; 15;7[12]:5511-6. eCollection 2014.
29. 30. Vardi N, Parlakpınar H, Ateş B, Çetin A, Otlu A. Antiapoptotic and antioxidant effects of beta carotene against methotrexate-induced testicular injury. Fertil. Steril, 2009; 92: 2028-33.
30. 31. Abud-Mendoza C, Martinez-Martinez MU, Monsivais-Urenda A, Gonzalez-Amaro R. Laboratory biomarkers for guiding therapy with methotrexate in rheumatoid arthritis. Curr Pharm Dess, 2014; 21[2]:202-11.
31. 32. Martin SA, McCarthy A, Barber LJ, Burgess DJ, Parry S, Lord CJ, Ashworth A. Methotrexate induces oxidative DNA damage and is selectively lethal to tumour cells with defects in the DNA mismatch repair gene MSH2. EMBO Mol Med, 2009;1[6-7]:323-37.
32. 33. Abbasi MM, Khiavi MM, Monfaredan A, Hamishehkar H, Seidi K, Jahanban-Esfahlan R. DOX-MTX-NPs augment p53 mRNA expression in OSCC model in rat: effects of IV and oral routes. Asiyan Pac J Cancer Prev.2014; 15[19]:8377-82.
33. 34. Kalender S, Uzun FG, Demir F, Uzunhisarcıklı M, Aslanturk A. Mercuric chloride-induced testicular toxicity in rats and the protective role of sodium selenite and vitamin E. Food Chem Toxicol, 2013; 55:456-62.
34. 35. Ayinde OC, Ogunnowo S, Ogedegbe RA. Influence of Vitamin C and Vitamin E on testicular zinc content and testicular toxicity in lead exposed albino rats. BMC Pharmacol Toxicol, 2012; 13:17.
35. 36. Chen H, Liu J, Luo L, Baig MU, Kim JM, Zirkin BR. Vitamin E, aging and Leydig cell steroido genesis. Exp Gerontol, 2005; 40[8-9]: 728-36.
36. 37. Shrestha S, Dhungel S, Saxena AK, Bhattacharya S, Maskey D. Effect of methotrexate [MTX] administration on spermatogenesis: an experimental on animal model. Nepal Med Coll J. 2007; 9[4]:230-3. 38. Nouri HS, Azarmi Y, Movahedin M. Effect of growth hormone on testicular dysfunction induced by methotrexate in rats. Androloqia; 2009; 41[2]:105-10. 39. Gökçe A, Oktar S, Koç A, Yonden Z. Protective effects of thymoquinone against methotrexate-induced testicular injury. Human and Experimental Toxicology 2011; 30[8] 897-903.
37. 40. Saxena AK, Dhungel S, Bhattacharya S, JHA CB, Srıvastava AK. Effect Of Chronic Low Dose Of Methotrexate On Cellular Proliferation During Spermatogenesis In Rats. Archives of Andrology 2004; 50: 33-35.
38. 41. Shirpoor A, Norouzi L, Khadem-Ansari MH, Ilkhanizadeh B, Karimipour M [2014]. The Protective Effect of Vitamin E on Morphological and Biochemical Alteration Induced by Pre and Postnatal Ethanol Administration in the Testis of Male Rat Offspring: A Three Months Follow-up Study. J Reprod Infertil 2014; 15[3]:134-41.
39. 42. Armagan A, Uzar E, Uz E, Yilmaz HR, Kutluhan S, Koyuncuoglu HR, Soyupek S, Cam H, Serel TA [2008]. Caffeic acid phenethyl ester modulates methotrexate-induced oxidative stress in testes of rat. Hum Exp Toxicol.2008; 27[7]: 547-52.
40. 43. Yuncu M, Bükücü N, Bayat N, Sencar L, Tarakçioğlu M. The effect of vitamin E and L-carnitine against methotrexate-induced injury in rat testis. Turk J Med Sci. 2015; 45[3]:517-25.