Galeati G, Giaretta E, Zannoni A, Bucci D, Tamanini C, Forni M, Spinaci M
Dipartimento di Scienze Mediche Veterinarie (DIMEVET), Universita di Bologna, Bologna, Italia.
J Physiol Pharmacol. 2016 Aug;67(4):513-519.
Oxidative stress caused from in vitro culture contributes to inadequate oocyte maturation which leads to a poor embryo development. Therefore, it is important to protect oocytes and embryos against oxidative stress. This study was aimed at evaluating the effect of Embelin (2,5-dihydroxy-3-undecyl-1,4-benzoquinone), an antioxidant with various pharmacologic activities, on nuclear and cytoplasmic maturation of pig oocytes as well as on steroidogenesis of cumulus cells (CCs). Another objective was to determine the influence of Embelin on developmental competence of pig oocytes as well as the expression levels of three key genes (Nanog, Sox2 and Oct4) involved in the control of pluripotency in parthenogenetically activated embryos. Embelin (0, 10, 20 and 40 μM) was added during in vitro maturation of cumulus oocyte complexes; media of both the first and the second day of culture were collected and assayed for progesterone and estradiol-17β. At the end of the maturation period, the oocytes were fixed (to determine nuclear maturation) or partenogenically activated to evaluate cytoplasmic maturation and genes expression. Embelin did not exert any effect on the proportion of MII oocytes, steroidogenesis of CCs, percentage of embryos that developed to blastocyst stage and the number of blastomeres/blastocyst. Moreover, no significant differences of Oct4, Nanog and Sox2 transcripts were detected in blastocyst stage embryos. In conclusion, Embelin did not influence the reproductive parameters assessed, confirming that it is not possible to predict whether the beneficial effect exerted by an antioxidant in a particular tissue could be present also in another one.
体外培养引起的氧化应激会导致卵母细胞成熟不足,进而导致胚胎发育不良。因此,保护卵母细胞和胚胎免受氧化应激的影响非常重要。本研究旨在评估具有多种药理活性的抗氧化剂恩贝林(2,5 - 二羟基 - 3 - 十一烷基 - 1,4 - 苯醌)对猪卵母细胞核成熟和细胞质成熟以及对卵丘细胞(CCs)类固醇生成的影响。另一个目的是确定恩贝林对猪卵母细胞发育能力以及孤雌激活胚胎中参与多能性控制的三个关键基因(Nanog、Sox2和Oct4)表达水平的影响。在卵丘 - 卵母细胞复合体的体外成熟过程中添加恩贝林(0、10、20和40 μM);收集培养第一天和第二天的培养基,检测孕酮和雌二醇 - 17β。在成熟期末,固定卵母细胞(以确定核成熟)或进行孤雌激活以评估细胞质成熟和基因表达。恩贝林对MII期卵母细胞的比例、CCs的类固醇生成、发育到囊胚阶段的胚胎百分比以及囊胚中的卵裂球数量均无影响。此外,在囊胚期胚胎中未检测到Oct4、Nanog和Sox2转录本的显著差异。总之,恩贝林不影响所评估的生殖参数,证实无法预测抗氧化剂在特定组织中发挥的有益作用是否也会在另一个组织中出现。