Masoumeh ZIRAK JAVANMARD, Mojtaba KARIMIPOUR, Bagher POURHEYDAR, Farideh DANESHPOYA

Effects of n-acetylcysteine on ovarian tissue autografted intogranulation tissue compared to back muscle in rats

Background/aim: Ovarian transplantation can preserve fertility in cancer patients. The aim of this study was to compare ovarian transplantation into granulation tissue (GT) versus back muscle (BM) sites and also to investigate the effects of N-acetylcysteine (NAC) as an antioxidant on ovarian survival in a rat autograft model. Materials and methods: Twenty-eight adult female rats were divided into four equal groups (n = 7 each) as follows: Group 1, ovarian tissue grafted into GT + saline (GT + saline); Group 2, ovarian tissue grafted into GT + NAC (GT + NAC); Group 3, ovarian tissue grafted into BM + saline (BM + saline); and Group 4, ovarian tissue grafted into BM + NAC (BM + NAC). After 28 days, serum concentrations of malondialdehyde (MDA), progesterone, and estradiol (E2) were measured. Ovarian follicle counts were performed from hematoxylin and eosin-stained slides. Furthermore, apoptosis was assessed by terminal deoxynucleotidyl transferase (TdT) dUTP nick-end labeling (TUNEL). Results: There were no significant differences found in levels of MDA, progesterone, and E2 among the groups. The percentage of apoptotic granulosa cells was significantly (P < 0.05) lower in the ovarian tissue autografted in the GT + NAC group compared to the GT + saline and BM + NAC groups. In addition, the number of blood vessels formed in the ovarian tissue in the GT + NAC group was significantly higher than in the GT + saline and BM + NAC groups (P < 0.05 and P < 0.001 respectively). Conclusion: The granulation tissue site is a suitable candidate for ovarian graft in rats. Furthermore, NAC could not restore autografted ovarian functions.

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1. Kim SS. Time to re-think: ovarian tissue transplantation versus whole ovary transplantation. Reprod Biomed Online 2010; 20: 171-174.
2. Dath C, Van Eyck A, Dolmans MM, Romeu L, Delle Vigne L, Donnez J, Van Langendonckt A. Xenotransplantation of human ovarian tissue to nude mice: comparison between four grafting sites. Hum Reprod 2010; 25: 1734-1743.
3. Mahmoodi M, Soleimani Mehranjani M, Shariatzadeh SM, Eimani H, Shahverdi A. Effects of erythropoietin on ischemia, follicular survival, and ovarian function in ovarian grafts. Reproduction 2014; 147: 733-741.
4. Soleimani R, Heytens E, Oktay K. Enhancement of neoangiogenesis and follicle survival by sphingosine-1- phosphate in human ovarian tissue xenotransplants. PLoS One 2011; 6: e19475.
5. Demeestere I, Simon P, Emiliani S, Delbaere A, Englert Y. Orthotopic and heterotopic ovarian tissue transplantation. Hum Reprod Update 2009; 15: 649-665.
6. Hemadi M, Abolhassani F, Akbari M, Sobhani A, Pasbakhsh P, Ahrlund-Richter L, Modaresi MH, Salehnia M. Melatonin promotes the cumulus-oocyte complexes quality of vitrifiedthawed murine ovaries; with increased mean number of follicles survival and ovary size following heterotopic transplantation. Eur J Pharmacol 2009; 618: 84-90.
7. Gao JM, Yan J, Li R, Li M, Yan LY, Wang TR, Zhao HC, Zhao Y, Yu Y, Qiao J. Improvement in the quality of heterotopic allotransplanted mouse ovarian tissues with basic fibroblast growth factor and fibrin hydrogel. Hum Reprod 2013; 28: 2784-2793.
8. Yang H, Lee HH, Lee HC, Ko DS, Kim SS. Assessment of vascular endothelial growth factor expression and apoptosis in the ovarian graft: can exogenous gonadotropin promote angiogenesis after ovarian transplantation? Fertil Steril 2008; 90: 1550-1558.
9. Dissen G, Lara H, Fahrenbach W, Costa M, Ojeda S. Immature rat ovaries become revascularized rapidly after autotransplantation and show a gonadotropin-dependent increase in angiogenic factor gene expression. Endocrinology 1994; 134: 1146-1154.
10. Oktay K, Türkçüoğlu I, Rodriguez-Wallberg KA. Four spontaneous pregnancies and three live births following subcutaneous transplantation of frozen banked ovarian tissue: what is the explanation? Fertil Steril 2011; 95: 804-807.
11. Soleimani R, Van der Elst J, Heytens E, Van den Broecke R, Gerris J, Dhont M, Cuvelier C, De Sutter P. Back muscle as a promising site for ovarian tissue transplantation, an animal model. Hum Reprod 2008; 23: 619-626.
12. Damous LL, Nakamuta JS, Carvalho AE, Carvalho KC, SoaresJr JM, Simões Mde J, Krieger JE, Baracat EC. Does adipose tissue-derived stem cell therapy improve graft quality in freshly grafted ovaries? Reprod Biol Endocrinol 2015; 13: 108-119.
13. Oktay K, Newton H, Mullan J, Gosden RG. Development of human primordial follicles to antral stages in SCID/hpg mice stimulated with follicle stimulating hormone. Hum Reprod 1998; 13: 1133-1138.
14. Israely T, Nevo N, Harmelin A, Neeman M, Tsafriri A. Reducing ischaemic damage in rodent ovarian xenografts transplanted into granulation tissue. Hum Reprod 2006; 21: 1368-1379.
15. Youm HW, Lee JR, Lee J, Jee BC, Suh CS, Kim SH. Transplantation of mouse ovarian tissue: Comparison of the transplantation sites. Theriogenology 2015; 83: 854-861.
16. Abdel-Ghani MA, Abe Y, Asano T, Suzuki H. Effect of graft site and gonadotrophin treatment on follicular development of canine ovarian grafts transplanted to NOD-SCID mice. Reprod Medand Biol 2011; 10: 259-266.
17. Lingen MW. Role of leukocytes and endothelial cells in the development of angiogenesis in inflammation and wound healing. Arch Pathol Lab Med 2001; 125: 67-71.
18. Abramovitch R, Marikovsky M, Meir G, Neeman M. Stimulation of tumour angiogenesis by proximal wounds: spatial and temporal analysis by MRI. Br J Cancer 1998; 77: 440-447.
19. Abir R, Fisch B, Jessel S, Felz C, Ben-Haroush A, Orvieto R. Improving posttransplantation survival of human ovarian tissue by treating the host and graft. Fertil Steril 2011; 95: 1205- 1210.
20. Li SH, Hwu YM, Lu CH, Chang HH, Hsieh CE, Lee RK. VEGF and FGF2 improve revascularization, survival, and oocyte quality of cryopreserved, subcutaneously-transplanted mouse ovarian tissues. Int J Mol Sci 2016; 17: 1-13.
21. Cuzzocrea S, Mazzon E, Costantino G, Serraino I, De Sarro A, Caputi AP. Effects of N-acetylcysteine in a rat model of ischemia and reperfusion injury. Cardiovasc Res 2000; 47: 537- 548.
22. Sener G, Tosun O, Kaçmaz A, Arbak S, Ersoy Y, AyanoğluDülger G. Melatonin and N-acetylcysteine have beneficial effects during hepatic ischemia and reperfusion. Life Sci 2003; 72: 2707-2718.
23. Sehirli AO, Sener G, Satiroglu H, Ayanoglu-Dulger G. Protective effect of N-acetylcysteine on renal ischemia/ reperfusion injury in the rat. J Nephrol 2003; 16: 75-80.
24. Ozdulger A, Cinel I, Koksel O, Cinel L, Avlan D, Unlu A, Okcu H, Dikmengil M, Oral U. The protective effect of N-acetylcysteine on apoptotic lung injury in cecal ligation and puncture-induced sepsis model. Shock 2003 ; 19: 366-372.
25. Mahmoodi M, Mehranjani MS, Shariatzadeh SMA, Eimani H, Shahverdi A. N-Acetylcysteine improves function and follicular survival in mice ovarian grafts through inhibition of oxidative stress. Reprod Biomed Online 2015; 30: 101-110.
26. Nasr A. Effect of N-acetyl-cysteine after ovarian drilling in clomiphene citrate-resistant PCOS women: a pilot study. Reprod Biomed Online 2010; 20: 403-409.
27. Zafarullah M, Li W, Sylvester J, Ahmad M. Molecular mechanisms of N-acetylcysteine actions. Cell Mol Life Sci 2003; 60: 6-20.
28. Amorim EM, Damous LL, Durando MC, Saraiva MV, Koike MK, Montero EF. N-Acetylcysteine improves morphologic and functional aspects of ovarian grafts in rats. Acta Cir Bras 2014; 29: 22-27.
29. Inci I, Zhai W, Arni S, Hillinger S, Vogt P, Weder W. N-Acetylcysteine attenuates lung ischemiareperfusion injury after lung transplantation. Ann Thorac Surg 2007; 84: 240-246.
30. Byers SL, Wiles MV, Dunn SL, Taft RA. Mouse estrous cycle identification tool and images. PLoS One 2012; 7: e35538.
31. Huang Y, Zhao X, Zhang Q, Xin X. The GnRH antagonist reduces chemotherapy-induced ovarian damage in rats by suppressing the apoptosis. Gynecol Oncol 2009; 112: 409-414.
32. Liu J, Van der Elst J, Van den Broecke R, Dhont M. Early massive follicle loss and apoptosis in heterotopically grafted newborn mouse ovaries. Hum Reprod 2002; 17: 605-611.
33. Yang HY, Cox SL, Jenkin G, Findlay J, Trounson A, Shaw J. Graft site and gonadotrophin stimulation influences the number and quality of oocytes from murine ovarian tissue grafts. Reproduction 2006; 131: 851-859.
34. Israely T, Dafni H, Granot D, Nevo N, Tsafriri A, Neeman M. Vascular remodeling and angiogenesis in ectopic ovarian transplants: a crucial role of pericytes and vascular smooth muscle cells in maintenance of ovarian grafts. Biol Reprod 2003; 68: 2055-2064.
35. Callejo J, Vilaseca S, Ordi J, Cabré S, Lailla JM, Balasch J. Heterotopic ovarian transplantation without vascular pedicle in syngeneic Lewis rats: long-term evaluation of effects on ovarian structure and function. Fertil Steril 2002; 77: 396-402.
36. Khan M, Sekhon B, Jatana M, Giri S, Gilg AG, Sekhon C, Singh I, Singh AK. Administration of N‐acetylcysteine after focal cerebral ischemia protects brain and reduces inflammation in a rat model of experimental stroke. J Neurosci Res 2004; 76: 519-527.