RESVERATROL SIÇANLARDA OKSİDATİF STRESİ VE APOPTOZU BASKILAYARAK METOTREKSAT KAYNAKLI YUMURTALIK HASARINI HAFİFLETİR

Year 2022, Volume: 29 Issue: 3, 476 - 484, 30.09.2022

Kübra Kısmet Aka , Çiğdem Aydın Acar , Meltem Özgöçmen , Rahime Aslankoç , Şükriye Yeşilot

https://doi.org/10.17343/sdutfd.1145034

Abstract

Amaç
Dişi sıçanlarda Metotreksat (MTX) kaynaklı yumurtalık
hasarına karşı Resveratrolün (RES) antioksidan ve
antiapoptotik etkilerinin değerlendirilmesi amaçlanmaktadır.
Gereç ve Yöntem
Çalışmada ratlar 7 gruba ayrıldı: Kontrol; MTX (15mg/
kg-1. gün); MTX (1. ve 3. gün); MTX (1., 3. ve 5. gün);
MTX (1. gün) + RES (20mg/kg); MTX (1. ve 3. gün)
+ RES; MTX (1., 3. ve 5. gün) + RES. Grup 4 ve 6'da
sadece bir hayvan kaldığı ve grup 7'de hiç hayvan
olmadığı için bu gruplar değerlendirme dışı bırakıldı.
Yumurtalık hasarı, sıçanların yumurtalık dokularında
hematoksilen-eozin (H-E) ve TUNEL boyaması yapılarak
değerlendirildi. Yumurtalık dokularındaki toplam
oksidan/antioksidan (TOS/TAS) durumu spektrofotometrik
olarak değerlendirildi. Ayrıca kandaki DNA hasarı
comet testi ile değerlendirildi.
Bulgular
Biyokimyasal analizler sonucunda MTX gruplarında,
grup 2 ve 3'te kontrol grubuna göre TOS düzeyleri artarken,
grup 5'te anlamlı olarak azaldı. Öte yandan,
grup 5'te grup 2 ve 3'e göre TAS düzeyi anlamlı olarak
arttı. H-E boyama sonuçları, MTX kaynaklı hasarın
RES (grup 5) tedavisi ile düzeldiğini gösterirken;
maksimum TUNEL pozitif boyama görüntüsü; 2. ve 3.
gruplarda eşit miktarlarda, ardından 5. grupta gözlemlenmiştir.
Ayrıca DNA hasarını gösteren comet skoru
2. ve 3. gruplarda anlamlı olarak artarken, 5. grupta
önemli ölçüde azalmıştır.
Sonuç
Bu çalışmanın sonuçları doğrultusunda, Resveratrol’ün
antioksidan özelliğinden dolayı Metotreksat’a
bağlı yumurtalık hasarına karşı koruyucu etki sağlayabileceği
görülmüştür.

Keywords

Metotreksat, Resveratrol, yumurtalık hasarı, antioksidan, rat

Supporting Institution

Burdur Mehmet Akif Ersoy Üniversitesi Bilimsel Araştırma Projeleri Koordinatörlüğü

Project Number

0615-YL-19

RESVERATROL ALLEVIATES METHOTREXATE-INDUCED OVARIAN INJURY VIA SUPPRESSING OXIDATIVE STRESS AND APOPTOSIS IN RATS

Abstract

Objective
The study aimed to evaluate the antioxidant and
antiapoptotic effects of Resveratrol (RES) against
Methotrexate (MTX)-induced ovarian damage in
female rats and were divided into 7 groups:
Material and Method
Control; MTX (15mg/kg-1st day); MTX (1st and 3rd
day); MTX (1st, 3rd and 5th day); MTX (1st day) +
RES (20mg/kg); MTX (1st and 3rd day) + RES; MTX
(1st, 3rd and 5th day) + RES. Since there was only
one animal left in groups 4 and 6 and no animals
in group 7, these groups were excluded from the
evaluation. Ovarian damage was evaluated by
performing hematoxylin-eosin (H-E) and TUNEL
staining on ovarian tissues of rats. Total oxidant/
antioxidant status in ovarian tissues was evaluated
spectrophotometrically. In addition, DNA damage in
blood was assessed by comet assay.
Results
As a result of biochemical analyzes, TOS levels
increased in MTX groups, group 2 and 3, compared
to the control group, while TOS level decreased
significantly in group 5. On the other hand, TAS
level increased significantly in group 5 compared
to groups 2 and 3. H-E staining results showed that
MTX-induced damage improved with RES (group 5)
treatment. Maximum TUNEL positive staining image;
it was observed in equal amounts in groups 2 and
3, followed by group 5. In addition, the comet score,
which indicates DNA damage, increased significantly
in groups 2 and 3, while it decreased significantly in
group 5.
Conclusion
In line with the results of this study, it was observed
that Resveratrol may provide protective effects
against Methotrexate-induced ovarian damage due to
its antioxidant properties.

Keywords

Methotrexate, Resveratrol, ovarian damage, rat, antioxidant

Project Number

0615-YL-19

References

• 1. Fridlington JL, Tripple JW, Reichenberg JS, Hall CS, Diven DG. Acute methotrexate toxicity seen as plaque psoriasis ulceration and necrosis: a diagnostic clue. Dermatol Online J 2011;17(11):2.
• 2. Hyoun SC, Običan SG, Scialli AR. Teratogen update: methotrexate. Birth Defects Res A Clin Mol Teratol 2012;94(4):187-207.
• 3. Karri S, Vanithakumari G. Lipid Alterations Instigated by Methotrexate and Leucovorin in Female Reproductive Tract of Albino Rats. International Journal of Cancer Research and Prevention 2011;4(3):253.
• 4. Widemann BC, Adamson PC. Understanding and managing methotrexate nephrotoxicity. Oncologist. 2006;11(6):694-703.
• 5. Kivity S, Zafrir Y, Loebstein R, Pauzner R, Mouallem M, Mayan H. Clinical characteristics and risk factors for low dose methotrexate toxicity: a cohort of 28 patients. Autoimmun Rev 2014;13(11):1109-1113.
• 6. Caron JE, Krull KR, Hockenberry M, Jain N, Kaemingk K, Moore IM. Oxidative stress and executive function in children receiving chemotherapy for acute lymphoblastic leukemia. Pediatr Blood Cancer 2009;53(4):551-556.
• 7. Oktem F, Yılmaz R, Ozguner F, Olgar S, Ayata A, Uzar E, Uz E. Methotrexate-induced renal oxidative stress in rats: the role of a novel antioxidant caffeic acid phenethyl ester. Toxicol Ind Health 2006;22(6):241-247.
• 8. Uygur AG, Shingirik E, Yucel AH. Protective effect of alpha lipoic acid in methotrexate-induced damage. Archive Rel J 2019;28(3):227-236.
• 9. Kabakci M, Kurutas EB, Bakaris S, Gungor M. The effect of erythropoietin on methotrexate-ınduced renal damage in rats: biochemical and histopathological studies. Çukurova Med J 2015;40(1):98-106.
• 10. Baxter RA. Anti-aging properties of resveratrol: review and report of a potent new antioxidant skin care formulation. J Cosmet Dermatol 2008;7(1):2-7.
• 11. Mazza G. Anthocyanins in grapes and grape products. Crit Rev Food Sci Nutr 1995; 35(4):341-371.
• 12. Karabulut AB. Resveratrol and its effects. Turkey Clinics J Med Sci 2008;28(6):166-169.
• 13. Keskin N, Noyan T, Kunter B. Health from Grape by Resveratrol: Review. J Med Sci 2009;29(5):1273-1279.
• 14. Mahady GB, Pendland SL. Resveratrol inhibits the growth of helicobacter pylori in vitro. Am J Gastroenterol 2000;95(7):1849.
• 15. Ortega I, Duleba AJ. Ovarian actions of resveratrol. Ann N Y Acad Sci 2015;1348(1):86-96.
• 16. Rauf A, Imran M, Butt MS, Nadeem M, Peters DG, Mubarak MS. Resveratrol as an anti-cancer agent: a review. Crit Rev Food Sci Nutr 2018;58(9):1428-1447.
• 17. Refaiy A, Muhammad E, ElGanainy EO. Semiquantitative smoothelin expression in detection of muscle invasion in transurethral resection and cystectomy specimens in cases of urinary bladder carcinoma. African J Urol 2011;17(1):6-10.
• 18. Chen CY, Wang YF, Huang WR, Huang YT. Nickel induces oxidative stress and genotoxicity in human lymphocytes. Toxicol Appl Pharmacol 2003;189(3):153-9.
• 19. Bausinger J, Speit G. The impact of lymphocyte isolation on induced DNA damage in human blood samples measured by the comet assay. Mutagenesis 2016;31(5):567-72.
• 20. Collins AR. Measuring oxidative damage to DNA and its repair with the comet assay. Biochim Biophys Acta 2014;1840(2):794-800.
• 21. Gradishar WJ, Schilsky RL. Effects of cancer treatment on the reproductive system. Critical Reviews in Oncology/Hematology 1988;8(2): 153-171.
• 22. Yulug E, Turedi S, Alver A, Turedi S, Kahraman C. Effects of resveratrol on methotrexate-induced testicular damage in rats. Scientific World Journal 2013;2013:489659.
• 23. Sonmez MF, Cilenk KT, Karabulut D, Unalmıs S, Deligonuul E, Oztuurk I, Kaymak E. Protective effects of propolis on methotrexate-induced testis injury in rat. Biomedicine & Pharmacotherapy 2016;79: 44-51.
• 24. Asci H, Ozmen O, Ellidag HY, Aydin B, Bas E, Yilmaz N. The impact of gallic acid on the methotrexate-induced kidney damage in rats. J Food Drug Anal 2017;25(4):890-897.
• 25. Erdogan E, Ilgaz Y, Gurgor PN, Oztas Y, Topal T, Oztas E. Rutin ameliorates methotrexate induced hepatic injury in rats1. Acta Cir Bras 2015;30(11):778-784.
• 26. Vardi N, Parlakpinar H, Cetin A, Erdogan A, Cetin Ozturk I. Protective effect of β-carotene on methotrexate–induced oxidative liver damage. Toxicol Pathol 2010;38(4):592-597.
• 27. Agarwal A, Aponte-Mellado A, Premkumar BJ, Shaman A, Gupta S. The effects of oxidative stress on female reproduction: a review. Reproductive Biology and Endocrinology 2012;10: 49.
• 28. Gusman J, Malonne H, Atassi G. A reappraisal of the potential chemopreventive and chemotherapeutic properties of resveratrol. Carcinogenesis 2001;22(8):1111-1117.
• 29. Tunali-Akbay T, Sehirli O, Ercan F, Sener G. Resveratrol protects against methotrexate-induced hepatic injury in rats. J Pharm Pharm Sci 2010;13(2):303-10.
• 30. Yulug E, Turedi S, Alver A, Turedi S, Kahraman C. Effects of resveratrol on methotrexate-induced intestinal injury. Bratisl Lek Listy 2015;116(11):676-80.
• 31. Arslan A, Ozcicek F, Keskin Cimen F, Altuner D, Yarali O, Kurt N, Tumkaya L, Ozturk C, Suleyman H. Protective effect of resveratrol against methotrexate-induced oxidative stress in the small intestinal tissues of rats. Int J Clin Exp Med 2015;8(7):10491-500.
• 32. Ozcicek A, Cetin N, Keskin Cimen F, Tumkaya L, Malkoc I, Gulaboglu M, Yarali O, Suleyman B. The Impact of Resveratrol on Oxidative Stress Induced by Methotrexate in Rat Ileum Tissue: Evaluation of Biochemical and Histopathological Features and Analysis of Gene Expression. Med Princ Pract 2016;25(2):181-6.
• 33. Senol E. What happened in febrile neutropenia in the last year? Factors and epidemiology (literature review). Third Febrile Neutropenia Postgraduate Training Course. February 27–29. Bilkent Hotel, Ankara 2004. https://www.febrilnotropeni.net/pdf/3_MSEK/literatur_degerlenensi.pdf (Access Date: 17.08.2021)
• 34. Gonderen HHS, Kapucu S. Evaluation criteria for neutropenia and nursing care in the neutropenic patient. Hacettepe University nursing faculty J 2009;16(1):69-75.
• 35. Kiremitli T, Kiremitli S, Erel O, Oguz E, Dinc K, Nayki U, Nayki C, Turkler C, Kirkinci A. Dynamic thiol/disulphide homeostasis and ischemic modified albumin levels in isolated oligohydramnios. Taiwan J Obstet Gynecol 2021;60(6):1038-1042
• 36. Ata N, Kulhan NG, Kulhan M, Turkler C, Kiremitli T, Kiremitli S, Keskin Cimen F, Suleyman H, Toprak V. The effect of resveratrol on oxidative ovary-damage induced by methotrexate in rats (Resveratrol oxidative ovary-damage). Gen Physiol Biophys 2019;38(6):519-524.
• 37. Turkler C, Onat T, Yildirim E, Kaplan S, Yazici GN, Mammadov R, Sunar M. An experimental study on the use of lycopene to prevent infertility due to acute oxidative ovarian damage caused by a single high dose of methotrexate. Adv Clin Exp Med 2020;29(1):5-11.
• 38. Soylu Karapinar O, Pinar N, Özcan O, Özgür T, Dolapçıoğlu K. Protective effect of alpha-lipoic acid in methotrexate-induced ovarian oxidative injury and decreased ovarian reserve in rats. Gynecol Endocrinol 2017;33(8):653-659.
• 39. Chen ZG, Luo LL, Xu, JJ, Zhuang XL, Kong XX, Fu YC. Effects of plant polyphenols on ovarian follicular reserve in aging rats. Biochem Cell Biol 2010;88(4):737-745.
• 40. Ozdemir HU. Investigation of the effects of resveratrol on cyclophosphamide-induced ovarian damage in rats at light and electron microscopic level. Istanbul University, Institute of Health Sciences, Istanbul 2014.
• 41. Gunyeli I, Saygin M, Ozmen O. Methotrexate-induced toxic effects and the ameliorating effects of astaxanthin on genitourinary tissues in a female rat model. Arch Gynecol Obstet 2021;304(4):985-997.
• 42. Yuksel Y, Yuksel R, Yagmurca M, Haltas H, Erdamar H, Toktas M, Ozcan O. Effects of quercetin on methotrexate-induced nephrotoxicity in rats. Hum Exp Toxicol 2017;36(1):51-61.
• 43. El-Sheikh AA, Morsy MA, Abdalla AM, Hamouda AH, Alhaider IA. Mechanisms of Thymoquinone Hepatorenal Protection in Methotrexate-Induced Toxicity in Rats. Mediators Inflamm 2015;2015:859383.
• 44. 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 toxic- ity in rats. Int J Clin Exp Med 2014;7:5551–5516
• 45. Alturfan AA, Tozan-Beceren A, Sehirli AO, Demiralp E, Sener G, Omurtag GZ. Resveratrol ameliorates oxidative DNA damage and protects against acrylamide-induced oxidative stress in rats. Mol Biol Rep 2012;39(4):4589-4596.