N-acetylcysteine improves morphologic and functional aspects of ovarian grafts in rats

Abstract

PURPOSE:To evaluate morphological and functional aspects of the ovarian graft in transplanted rats treated with NAC. METHODS:Female Wistar rats, virgin, 3 to 4 months old, weighing 200-250 grams were used in experiments. The rats have been kept in proper sanitary conditions, receiving food and water ad libitum. Five groups (n=10, each) were constituted: 4 groups treated subcutaneously with NAC, at doses of 150, 300, 600 and 1200 mg/kg (NAC150, NAC300, NAC600 and NAC1200, respectively), one hour of before the ovarian transplantation and control group (GTx) -treated with physiological solution and submitted to ovarian transplantation. The rats were anesthetized and submitted to autologous left ovarian transplantation, without anastomosis in retroperitoneum, and contralateral oophorectomy. During follow-up of 4 or 15 days, the estrous cycle was evaluated by vaginal smears to determine cycle regularity. At the end of 4 th or 15 th days, rats were re-anesthetized and blood and graft were obtained to estradiol analysis and morphological assessment. Data were analysed by One Way Analysis of Variance (ANOVA) or ANOVA on ranks complemented by Student-Newman-Keuls test. RESULTS:At 4 th day, viable follicles in the graft did not altered by NAC treatments. The NAC300 and NAC600 groups showed increasing in follicle atresia (p=0.012) compared to GTx and NAC1200 group. At 15 th day, 50% of GTx, NAC150, and NAC300 rats showed regular oestrous cycle; 83% of NAC600 and 100% of NAC1200 rats returned to regular cycle. NAC1200 group showed increasing in primordial follicle compared to GTx, NAC150 or NAC300 (p=0.011). NAC did not interfere in estradiol levels after 4 or 15 days of transplantation. CONCLUSION:In autologous ovarian transplantation, high dose of NAC promotes graft viability with recovery of estrous cycle.

Remote Ischemic Preconditioning and N-Acetylcysteine in Autologous Ovarian Transplantation in Rats.

“…Likewise, the early application of N-acetylcysteine (NAC) is related to beneficial effects in ovarian graft preservation [11]. This substance is a tiol of low molecular weight derivated of cysteine and acts removing reactive oxygen species, in addition to improving nitric oxide activity and regulating microcirculation [12].…”

“…On 15 th postoperative day [11], animals were euthanized with a lethal dose of previously used anesthetics.…”

“…One hour before ovarian transplantation, animals were treated subcutaneously -along the dorsum -with 1200mg/kg of NAC [11]. The non-treated groups, namely controls, GTx and R-IPC were treated with 0.9% Sodium chloride solution at the same volume.…”

“…On the 15 ℎ post-operative day [11], blood samples were collected from the inferior vena cava for estradiol and progesterone serum levels (tested with kits in a solid-phase, competitive, imunofluorimetry and radioimmunoassay, respectively) Histopathology:-Ovarian grafts were surgically removed from the inguinal region and fixed in 10% formaldehyde. The sections were dewaxed and gradually hydrated before being stained with hematoxylin-eosin.…”

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

“…Mahmoodi et al showed that NAC improves follicular survival in mice ovarian autografts into the BM by decreasing oxidative stress and apoptosis (25). Amorim et al also reported that subcutaneous injection of NAC at a high dose improves follicular survival of ovarian tissue grafted into a retroperitoneal position (28).…”