Rat Overinde İskemi-Reperfüzyon Üzerine N-Asetil Sistein ve Resveratrol’ün Koruyucu Etkisi

Abstract

Amaç: Bu çalışmanın amacı, rat overinde iskemi-reperfüzyona bağlı doku hasarı üzerine N-asetil sistein ve resveratrol’ün etkisini değerlendirmektir. Yöntemler: 42 adet dişi Wistar rat temin edildi. Bu ratlar randomize bir şekilde 6 gruba eşit olarak bölündü (n=7). Oluşturulan gruplar; sham (S), torsiyon (T), torsiyon - detorsiyon (T - D), torsiyon - detorsiyon + salin (T - DT - S), torsiyon - detorsiyon + resveratrol (20 mg/kg) (T - DT - Res) ve torsiyon - detorsiyon + N-asetil sistein (150 mg/kg) (T - DT - NAC) şeklinde işlemlere tabi tutuldu. Sham grubu hariç diğer tüm gruplara iki saat boyunca ovaryan torsiyon işlemi uygulandı. Torsiyon grubu dışındaki diğer tüm gruplara 2 saat detorsiyon prosedürü uygulandı. Salin, resveratrol ve N-asetil sistein gruplarında yer alan ratlara; detorsiyon işleminden yarım saat önce intraperitoneal yoldan 2 ml serum fizyolojik, 20 mg/kg resveratrol ve 150 mg/ kg N-asetil sistein uygulandı. Ardından tüm ratlardan oksidatif stres markerları ve tümör nekrotizan faktör alfa (TNF-α) düzeylerinin çalışılması için 2 ml kan örneği ile histolojik inceleme için torsiyone edilen overler çıkarıldı. Histopatolojik değişiklikler ödem, konjesyon, hemoraji, lökosit infiltrasyonu ve follikül dejenerasyonu şeklinde tanımlandı. Bulgular: Histopatolojik hasar skorlamasına göre en az hasar sham grubunda, en fazla ise torsiyon-detorsiyon grubunda olduğu görüldü (1,00±0,81, 11,00±1,15, p<0,001, p<0,001, sırasıyla). Resveratrol ile N-asetil sistein tedavisinin doku hasarını azaltmada etkili olduğu (total hasar skoru ortalaması: (83,85±0,89 ve 3,85±0,89 sırasıyla; p<0,001), buna karşın her iki ilaç arasında histopatolojik hasarın azaltılmasında anlamlı bir farklılığın olmadığı görüldü (p=0,966). Torsiyon-detorsiyon grubunda oksidatif stres düzeylerinin daha yüksek olduğu, resveratrol ile N-asetil sistein tedavisinin ise oksidatif stres düzeylerinde belirgin bir azalmaya neden olduğu saptandı. Ayrıca TNF-alfa düzeylerinin ilaç verilen gruplarda anlamlı ölçüde azaldığı tespit edildi (7,85±2,08 ve 8,68±1,88 sırasıyla; p<0,001). Buna karşın TNF-α düzeylerini azaltmada her iki ilacın da eşit etkinlikte olduğu gözlendi (8,68±1,88 ve 7,85±2,08, sırasıyla; p=0,968). Sonuç: Resveratrol ve NAC tedavisinin ovaryan reperfüzyona bağlı gelişen oksidatif stres ve doku hasarının azaltılmasında etkili olabileceği düşünülmektedir. Her iki ilaç arasında koruyucu etkinlik açısından bir fark bulunmadı.

Keywords

• İskemi-reperfüzyon, over, N-asetilsistein, resveratrol

Protective Effect of N-Acetylcystein and Resveratrol on Ischemia-Reperfusion Injury in Rat Ovary

Abstract

Objective: The aim of this study is evaluating the protective activity of N-acetyl cysteine and resveratrol treatment against ischemia - reperfusion damage created experimentally in rat ovaries. Methods: 42 female Wistar rats were used in our study. Rats were separated randomly into six groups consisting of seven rats as sham, torsion, torsion- detorsion, torsion-detorsion+saline, torsion-detorsion+resveretrol (20 mg/kg) and torsion- detorsion+N-acetyl cysteine (150 mg/kg). Except Sham, ovarian torsion procedure was implemented to all other groups for 2 hours. Detorsion procedure was implemented to other groups for 2 hours, except the torsion group. Medications were given through intraperitoneal way half an hour before the detorsion procedure in saline (two milliliter), resveratrol (20 mg/kg) and N-acetyl cysteine (150 mg/kg) groups. Then, 2 ml of blood samples were drawn for markers of oxidative stress and tumour necrosis factor-alpha (TNF-α) work and the ovaries, which were torsioned for the histologic examination, were ex­tracted from all rats. Edema, congestion, hemorrhage, leuko­cyte infiltration and degeneration of follicles were evaluated by histopathological examination. Results: According to histopathologic damage scores, the least damage was seen in sham group and the most damage was seen T-DT group (1.00±0.81 vs. 11.00±1.15, respectively; p<0.001). It was seen that resveratrol and N-acetyl cysteine treatments were effective in decreasing tissue damage (total damage score average 83.85±0.89 vs. 3.85±0.89, respec­tively; p<0.001), and on the other hand there was not any dif­ference between resveratrol and N-acetyl cysteine treatments (p=0.966). Besides, it was determined that oxidative stress levels were higher in torsion - detorsion group and the resve­ratrol and N-acetyl cysteine treatment caused a significant de­crease in oxidative stress levels. In additionally, the reductions of TNF-α levels were found to be equally effective in both drugs (8.68±1.88 vs. 7.85±2.08, P=0.968). Conclusion: Presented study showed that resveratrol and N-acetyl cysteine treatment can be effective in preventing tissue damage and oxidative stress, which is induced by ischemia-reperfusion that is created in rat ovaries. On the other hand, no difference was found between the resveratrol and N-acetyl cysteine with regards to protective activity.

Keywords

• schemia-reperfusion, ovary, N-acetylcysteine, resveratrol

References

1. 1. Hibbard LT. Adnexal torsion. Am J Obstet Gynecol 1985;152:456-461.
2. 2. Mage G, Canis M, Mahnes H, et al. Laparoscopic management of adnexal torsion. A review of 35 cases. J Reprod Med 1989;34:520-524.
3. 3. Porpora MG, Gomel V. The role of laparoscopy in the management of pelvic pain in women of reproductive age. Fertil Steril 1997;68:765-779.
4. 4. Meyer JS, Harmon CM, Harty MP, et al. Ovarian torsion: Clinical and imaging presentation in children. J Pediatr Surg 1995;30:1433-1436.
5. 5. Celik O, Turkoz Y, Hascalik S, et al. The protective effect of caffeic acid phenethyl ester on ischemia-reperfusion injury in rat ovary. Eur J Obstet Gynecol Reprod Biol 2004;117:183-188.
6. 6. Cakir Gungor AN, Gencer M, Karaca T, et al. The effect of hesperetin on ischemia-reperfusion injury in rat ovary. Arch Gynecol Obstet 2014;290:763-769.
7. 7. Sahin FK, Cosar E, Koken G, et al. Protective effect of aprotinin on ischemia-reperfusion injury in rat ovary. J Obstet Gynaecol Res 2008;34:794-800.
8. 8. Ustundag UV, Sahin S, Ak K,et al. The effects of tacrolimus on the activity and expression of tissue factor in the rat ovary with ischemia-reperfusion induced injury. Reprod Biol 2015;15:139-145.

9. 9. Akdemir A, Sahin C, Erbas O, et al. Is ursodeoxycholic acid crucial for ischemia / reperfusion-induced ovarian injury in rat ovary? Arch Gynecol Obstet 2015;292:445-450.
10. 10. Soleas GJ, Diamandis EP, Goldberg DM. Wine as a biological fluid: history, production, and role in disease prevention. J Clin Lab Anal 1997;11:287-313.
11. 11. Cotgreave IA. N-acetylcysteine: pharmacological considerations and experimental and clinical applications. Adv Pharmacol 1997;38:205-227.
12. 12. Ergun Y, Koc A, Dolapcioglu K, et al. The protective effect of erythropoietin and dimethylsulfoxide on ischemia-reperfusion injury in rat ovary. Eur J Obstet Gynecol Reprod Biol 2010;152:186-190.
13. 13. Eser A, Hizli D, Haltas H, et al. Effects of curcumin on ovarian ischemia- reperfusion injury in a rat model. Biomed Rep 2015;3:807-813.
14. 14. Gedik E, Girgin S, Ozturk H, et al. Resveratrol attenuates oxidative stress and histological alterations induced by liver ischemia/reperfusion in rats. World J Gastroenterol 2008;14:7101- 7106.
15. 15. Incebiyik A, Seker A, Camuzcuoglu H, et al. Does sildenafil have protective effects against ovarian ischemia-reperfusion injury in rats? Arch Gynecol Obstet 2015;291:1283-1288.
16. 16. Abali R, Tasdemir N, Yuksel MA, et al. Protective effect of infliximab on ischemia/reperfusion injury in a rat ovary model: biochemical and histopathologic evaluation. Eur J Obstet Gynecol Reprod Biol 2013;171:353-357.
17. 17. Buyukhatipoglu H, Kirhan I, Vural M, et al. Oxidative stress increased in healthcare workers working 24-hour on-call shifts. Am J Med Sci 2010;340:462-467.
18. 18. Sayyah-Melli M, Rashidi MR, Kaseb-Ganeh M, et al. The effect of erythropoietin against oxidative damage associated with reperfusion following ovarian detorsion. Eur J Obstet Gynecol Reprod Biol 2012;162:182-186.
19. 19. Kara M, Daglioglu YK, Kuyucu Y, et al. The effect of edaravone on ischemia-reperfusion injury in rat ovary. Eur J Obstet Gynecol Reprod Biol 2012;162:197-202.
20. 20. Maretta M, Bujdos M, Toth S Jr, et al. Alterations of epithelial layer after ischemic preconditioning of small intestine in rats. J Mol Histol 2012;43:171-178.
21. 21. Bozkurt S, Arikan DC, Kurutas EB, et al. Selenium has a protective effect on ischemia/reperfusion injury in a rat ovary model: biochemical and histopathologic evaluation. J Pediatr Surg 2012; 47:1735-1741.
22. 22. Yılmaz H, Sahin S, Sayar N, et al. Effects of folic acid and Nacetylcysteine on plasma homocysteine levels and endothelial function in patients with coronary artery disease. Acta Cardiol 2007;62:579-585.
23. 23. Millea PJ. N-Acetylcysteine: Multiple clinical applications. Am Fam Physician 2009;80:265-269.
24. 24. Sun Z, Lasson A, Olanders K, et al. Gut barrier permeability, reticuloendothelial system function and protease inhibitor levels following intestinal ischaemia and reperfusion effects of pretreatment with N-acetyl-L-cysteine and indomethacin. Dig Liver Dis 2002;34:560-569.
25. 25. Cay A, Alver A, Kucuk M, et al. the effects of N-aceyylcystein on antioxidant enzyme activities in experimental testicular torsion. J Surg Res 2006;131:199-203.
26. 26. Smyrniotis V, Arkadopoulos N, Kostapanaqiotou G, et al. Attenuation of ischemic injury by N-acetylcysteine preconditioning of the liver. J Surg Res 2005;129:31-37.
27. 27. Demir S, Inal-Erden M. Pentoxifylline and N-acetylcysteine in hepatic ischemia/reperfusion injury. Clin Chim Acta 1998;275:127-135.
28. 28. Hoch JR, Stevens RP, Keller MP, et al. Recovery of neuromuscular function during reperfusion of the ischemic extremity: effect of mannitol and superoxide dismutase. Surgery 1991;110:656-662.
29. 29. Bhalodia Y, Kanzariya N, Patel R, et al. Renoprotective activity of benincasa cerifera fruit extract on ischemia/reperfusioninduced renal damage in rat. Iran J Kidney Dis 2009;3:80-85.
30. 30. Yuan GJ, Ma JC, Gong ZJ, et al. Modulation of liver oxidant-antioxidant system by ischemic preconditioning during ischemia/reperfusion injury in rats. World J Gastroenterol 2005;11:1825-1828.
31. 31. Huang SS, Tsai MC, Chih CL, et al. Resveratrol reduction of infarct size in Long-Evans rats subjected to focal cerebral ischemia. Life Sci 2001;69:1057-1065.
32. 32. Hassan-Khabbar S, Cottart CH, Wendum D, et al. Postischemic treatment by trans-resveratrol in rat liver ischemia-reperfusion: a possible strategy in liver surgery. Liver Transpl 2008;14:451- 459.