Faculdade de Medicina FMUSP, Universidade de Sao Paulo, Av. Dr. Arnaldo, 455 - Cerqueira César, São Paulo, SP, CEP 01246-903, Brazil.
Instituto de Ciencias Biomedicas ICB, Universidade de Sao Paulo, Av. Prof. Lineu Prestes, 1374 - Butantã, São Paulo, SP, CEP 05508-000, Brazil.
Reprod Biol Endocrinol. 2021 Feb 3;19(1):17. doi: 10.1186/s12958-021-00705-4.
Melatonin has anti-inflammatory and antioxidative actions at the mitochondrial level. This indole-containing molecule may protect ovarian grafts during the process of cryopreservation. Therefore, we aimed to determine whether melatonin pretreatment improves rat ovarian graft quality.
Twenty-six female rats were allocated to two study groups of thirteen animals each: 1) control group: ovaries cryopreserved using the standard protocol; and 2) melatonin group: ovaries cryopreserved in a medium with melatonin. Ten rats of each group were submitted to 24-h freezing, and whole ovaries autologous and avascular transplantation with retroperitoneal placement. After postoperative (PO) day 15, daily vaginal smears were obtained for estrous cycle characterization. Between PO days 30 and 35, the animals were euthanized and ovarian grafts were recovered for histological and immunohistochemical (Ki-67, cleaved caspase-3, TUNEL, von Willebrand factor, estrogen, and progesterone receptors) analyses. The ovaries of the three remaining rats from each group were studied immediately after thawing to assess the effects of cryopreservation. ANOVA and Tukey's tests were used and the rejection level of the null hypothesis was set at 0.05 or 5% (p < 0.05).
Melatonin promoted faster restart of the estrous cycle and increased the expression of mature follicles, collagen type I, von Willebrand factor, Ki-67, and cleaved caspase-3 on corpora lutea and estrogen receptors in the ovaries as compared to control. There was a reduction in apoptosis by TUNEL on follicles, corpora lutea, and collagen type III.
Based on the evaluated parameters, melatonin may promote the quality of ovarian grafts. Reproductive function enhancement should be further studied.
褪黑素在线粒体水平具有抗炎和抗氧化作用。这种含吲哚的分子可能在冷冻保存过程中保护卵巢移植物。因此，我们旨在确定褪黑素预处理是否能提高大鼠卵巢移植物的质量。
26 只雌性大鼠分为两组，每组 13 只：1）对照组：使用标准方案冷冻卵巢；2）褪黑素组：在含褪黑素的培养基中冷冻卵巢。每组 10 只大鼠接受 24 小时冷冻，进行自体和无血管卵巢移植，放置于腹膜后。术后第 15 天，每天采集阴道涂片以确定动情周期特征。术后第 30-35 天，处死动物，回收卵巢移植物进行组织学和免疫组织化学（Ki-67、cleaved caspase-3、TUNEL、血管性血友病因子、雌激素和孕激素受体）分析。每组的另外 3 只大鼠的卵巢在解冻后立即进行研究，以评估冷冻保存的影响。采用方差分析和 Tukey 检验，假设检验的拒绝水平设定为 0.05 或 5%（p<0.05）。
与对照组相比，褪黑素促进了动情周期更快地恢复，增加了黄体和卵巢中成熟卵泡、胶原 I 型、血管性血友病因子、Ki-67 和 cleaved caspase-3 的表达以及雌激素受体。TUNEL 检测到卵泡、黄体和胶原 III 型的凋亡减少。
基于评估的参数，褪黑素可能会提高卵巢移植物的质量。应进一步研究生殖功能增强。
