Saeed Musapoor, Najmeh Davoodian, Ali Kadivar, Ebrahim Ahmadi, Hassan Nazari
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Matured oocytes were divided into two parts: one evaluated for their nuclear maturation, the level of GSH and ROS, mitochondrial membrane potential (MMP) and the pattern of transcription in oocytes and respective cumulus cells (CCs), and another subjected to fertilisation and culture to assess the development of oocytes to the blastocyst.</p><p><strong>Results: </strong>γ-Oryzanol improved the proportion of cleaved embryos and total blastocysts in the treated group, which was linked to improved MMP, higher levels of intracellular GSH and lower levels of ROS. A lower proportion of MI and GVBD was recorded for treated oocytes in comparison with control, although the proportion of MII oocytes was not different between groups. The treated oocytes and CCs showed downregulation of genes related to apoptosis (BAX and CASP-9) and upregulation of genes related to antioxidative status (NRF2, CAT and SOD). 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引用次数: 0
摘要
背景:绵羊卵母细胞体外成熟过程尚未取得可接受的结果:本研究旨在评估γ-oryzanol作为成熟培养基的补充物对雌性卵母细胞发育成囊胚的影响:方法:从雌性卵巢中抽取形态正常的卵母细胞-卵母细胞复合体(COCs),在添加或不添加 5 µM γ-oryzanol 的培养基中进行成熟。成熟的卵母细胞分为两部分:一部分用于评估其核成熟度、GSH 和 ROS 水平、线粒体膜电位(MMP)以及卵母细胞和相关积层细胞(CCs)的转录模式;另一部分用于受精和培养,以评估卵母细胞发育到囊胚的情况。结果:γ-Oryzanol 提高了处理组中裂解胚胎的比例和囊胚总数,这与 MMP 的改善、细胞内 GSH 水平的提高和 ROS 水平的降低有关。与对照组相比,处理组卵母细胞的 MI 和 GVBD 比例较低,但各组间 MII 卵母细胞的比例没有差异。经处理的卵母细胞和 CC 显示与细胞凋亡相关的基因(BAX 和 CASP-9)下调,而与抗氧化状态相关的基因(NRF2、CAT 和 SOD)上调。总之,我们的研究结果表明,补充剂可改善卵母细胞的发育结果,从而维持抗氧化反应和较高的酶活性,并阻止 ROS 的产生;因此,γ-玉兰脂醇通过抗氧化途径为对抗绵羊卵母细胞成熟过程中的氧化应激提供了一种新的选择。
Media Supplementation With Gamma-Oryzanol Improves the Outcome of Ovine Oocyte Maturation In Vitro.
Background: The process of maturing ovine oocyte in vitro has not yet been raised with acceptable results.
Objective: This study was designed to evaluate the γ-oryzanol effect as a supplement of maturation media on the development of ovine oocytes to blastocyst.
Methods: Aspirated from ovine ovaries, morphologically normal cumulus-oocyte complexes (COCs) were matured in media supplemented with or without 5 µM γ-oryzanol. Matured oocytes were divided into two parts: one evaluated for their nuclear maturation, the level of GSH and ROS, mitochondrial membrane potential (MMP) and the pattern of transcription in oocytes and respective cumulus cells (CCs), and another subjected to fertilisation and culture to assess the development of oocytes to the blastocyst.
Results: γ-Oryzanol improved the proportion of cleaved embryos and total blastocysts in the treated group, which was linked to improved MMP, higher levels of intracellular GSH and lower levels of ROS. A lower proportion of MI and GVBD was recorded for treated oocytes in comparison with control, although the proportion of MII oocytes was not different between groups. The treated oocytes and CCs showed downregulation of genes related to apoptosis (BAX and CASP-9) and upregulation of genes related to antioxidative status (NRF2, CAT and SOD). In conclusion, our results demonstrated the improved developmental outcome of supplemented oocytes so that the antioxidant response and higher enzymatic activity were maintained, and the generation of ROS was turned off; therefore, a novel alternative for counteracting oxidative stress in ovine oocytes undergoing maturation was offered by γ-oryzanol through an antioxidative pathway.
期刊介绍:
Veterinary Medicine and Science is the peer-reviewed journal for rapid dissemination of research in all areas of veterinary medicine and science. The journal aims to serve the research community by providing a vehicle for authors wishing to publish interesting and high quality work in both fundamental and clinical veterinary medicine and science.
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