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甘氨酸处理通过抑制细胞凋亡来提高猪卵母细胞和早期胚胎的发育潜能。

Glycine treatment enhances developmental potential of porcine oocytes and early embryos by inhibiting apoptosis.

机构信息

College of Animal Science and Technology, Jilin Agricultural University, Changchun, China.

Jilin Provincial Key Laboratory of Transgenic Animal and Embryo Engineering, Yanbian University, Yanji, Jilin, China.

出版信息

J Anim Sci. 2018 Jun 4;96(6):2427-2437. doi: 10.1093/jas/sky154.

Abstract

Glycine, a component of glutathione (GSH), plays an important role in protection from reactive oxygen species (ROS) and inhibition of apoptosis. The aim of this study was to determine the effect of glycine on in vitro maturation (IVM) of porcine oocytes and their developmental competence after parthenogenetic activation (PA). We examined nuclear maturation, ROS levels, apoptosis, mitochondrial membrane potential (ΔΨm), and ATP concentration, as well as the expression of several genes related to oocyte maturation and development. Our studies found that treatment with glycine in IVM culture medium increased nuclear maturation rate, but varying the concentrations of glycine (0.6, 6, or 12 mM) had no significant effect. Furthermore, 6 mM glycine supported greater blastocyst formation rates and lesser apoptosis after PA than the other concentrations (P < 0.05). All the glycine treatment groups had decreased levels of ROS in both matured oocytes and at the 2-cell stage (P < 0.05). At the 2-cell stage, the 6 mM glycine group had ROS levels that were lesser than the other 2 glycine treatment groups (0.6 and 12 mM). From this, we deemed 6 mM to be the optimal condition, and we then investigated the effects of 6 mM glycine on gene expression. The expression of both FGFR2 and Hsf1 were greater than the control group in mature oocytes. The glycine treatment group had greater levels of expression of an antiapoptotic gene (Bcl2) in mature oocytes and cumulus cells and lesser levels of expression of a proapoptotic gene (Bax) in PA blastocysts (P < 0.05). In addition, mitochondrial ΔΨm and ATP concentration were increased in 6 mM glycine group compared with the control group. In conclusion, our results suggest that glycine plays an important role in oocyte maturation and later development by reducing ROS levels and increasing mitochondrial function to reduce apoptosis.

摘要

甘氨酸是谷胱甘肽(GSH)的组成部分,在保护活性氧(ROS)和抑制细胞凋亡方面发挥重要作用。本研究旨在确定甘氨酸对猪卵母细胞体外成熟(IVM)及其孤雌激活(PA)后发育能力的影响。我们检查了核成熟度、ROS 水平、凋亡、线粒体膜电位(ΔΨm)和 ATP 浓度,以及与卵母细胞成熟和发育相关的几个基因的表达。我们的研究发现,在 IVM 培养基中添加甘氨酸可提高核成熟率,但不同浓度的甘氨酸(0.6、6 或 12 mM)对核成熟率没有显著影响。此外,6 mM 甘氨酸支持比其他浓度更高的 PA 后囊胚形成率和更少的凋亡(P < 0.05)。所有甘氨酸处理组在成熟卵母细胞和 2 细胞阶段的 ROS 水平均降低(P < 0.05)。在 2 细胞阶段,6 mM 甘氨酸组的 ROS 水平低于其他 2 个甘氨酸处理组(0.6 和 12 mM)。我们认为 6 mM 是最佳条件,然后研究了 6 mM 甘氨酸对基因表达的影响。在成熟卵母细胞中,FGFR2 和 Hsf1 的表达均高于对照组。甘氨酸处理组的成熟卵母细胞和卵丘细胞中抗凋亡基因(Bcl2)的表达水平较高,而 PA 囊胚中促凋亡基因(Bax)的表达水平较低(P < 0.05)。此外,与对照组相比,6 mM 甘氨酸组的线粒体ΔΨm 和 ATP 浓度增加。总之,我们的研究结果表明,甘氨酸通过降低 ROS 水平和增加线粒体功能来减少凋亡,从而在卵母细胞成熟和随后的发育中发挥重要作用。

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