Laboratory of Manipulation of Oocytes and Preantral Follicles (LAMOFOPA), Faculty of Veterinary Medicine, State University of Ceara, Av Dr Silas Munguba, 1700, Campus do Itaperi, Fortaleza, Ceara, Brazil.
Department of Physiology, Veterinary Faculty, University of Murcia, Murcia, Spain.
Reprod Sci. 2020 Aug;27(8):1602-1608. doi: 10.1007/s43032-020-00190-x.
Oxidative stress is one of the most detrimental factors that affect oocyte developmental competence and embryo development in vitro. The impact of anethole supplementation to in vitro maturation (IVM) media on oocyte maturation and further bovine in vitro embryo production was investigated. Oocytes of slaughterhouse-derived bovine ovaries were placed in IVM with anethole at different concentrations of 30 (AN30), 300 (AN300), and 2000 μg/mL (AN2000), or without (control treatment). The oocytes were assessed for maturation rates, and for reactive oxygen species (ROS) and ferric reducing antioxidant power (FRAP) levels, and mitochondrial membrane potential. Embryo development was assessed by cleavage and blastocyst rates, and embryo cell number. The percentage of metaphase II oocytes were similar among the treatments (range, 77%-96%). Anethole at 300 μg/mL was the only treatment that yielded higher cleavage and embryo development (morula and blastocyst) rates compared to the control treatment. The ROS production in the oocytes after maturation did not differ among treatments. However, oocytes treated with anethole at 300 μg/mL had higher (P < .05) FRAP and mitochondrial membrane potential compared to the control treatment. Furthermore, AN300 treatment increased (P < .05) the average number of total cells in blastocysts compared to the control and AN30 treatments. The use of anethole at 300 μg/mL during IVM is suggested to improve the quantity and quality of bovine embryos produced in vitro. The beneficial effects of anethole on embryonic developmental competence in vitro seems to be related to its capacity to regulate the redox balance and improve mitochondrial function in oocytes and embryos.
氧化应激是影响卵母细胞体外发育能力和胚胎发育的最有害因素之一。本研究旨在探讨茴香脑（anethole）补充到体外成熟（IVM）培养基中对卵母细胞成熟以及进一步的牛体外胚胎生产的影响。屠宰场来源的牛卵巢卵母细胞在含有不同浓度茴香脑（30 微克/毫升（AN30）、300 微克/毫升（AN300）和 2000 微克/毫升（AN2000））或无茴香脑（对照处理）的 IVM 培养基中进行培养。评估卵母细胞的成熟率以及活性氧（ROS）和铁还原抗氧化能力（FRAP）水平和线粒体膜电位。通过卵裂和囊胚率以及胚胎细胞数量评估胚胎发育情况。在处理之间，中期 II 期卵母细胞的百分比相似（范围为 77％-96％）。与对照处理相比，只有茴香脑浓度为 300 微克/毫升的处理能产生更高的卵裂和胚胎发育（桑葚胚和囊胚）率。成熟后卵母细胞中的 ROS 产生在处理之间没有差异。然而，与对照处理相比，用 300 微克/毫升茴香脑处理的卵母细胞具有更高的 FRAP 和线粒体膜电位（P<.05）。此外，与对照和 AN30 处理相比，AN300 处理增加了囊胚中总细胞数的平均值（P<.05）。因此，建议在 IVM 中使用 300 微克/毫升的茴香脑来提高体外生产的牛胚胎的数量和质量。茴香脑对体外胚胎发育能力的有益影响似乎与其调节氧化还原平衡和改善卵母细胞和胚胎中线粒体功能的能力有关。
