Hao Yuchun, Wang Jiapeng, Ren Jingyu, Liu Zhanpeng, Bai Ziyi, Liu Gang, Dai Yanfeng
College of Life Science, Inner Mongolia University, 235 West Univ. Road, Hohhot, 010021, Inner Mongolia, China.
Key Laboratory of Medical Cell Biology, Clinical Medicine Research Center, Affiliated Hospital of Inner Mongolia Medical University, 1 Tongdao North Street, Hohhot, 010050, Inner Mongolia, China.
Theriogenology. 2022 May;184:171-184. doi: 10.1016/j.theriogenology.2022.03.013. Epub 2022 Mar 17.
The supplementation of dimethyl alpha-ketoglutarate (DMKG) during the in vitro maturation (IVM) process has been shown to improve the in vitro developmental competences of porcine oocytes. Here, the effects of DMKG supplementation in IVM medium on the development competencies of ovine oocytes were investigated by analyzing the nuclear maturation rate to metaphase II (MII) stage, ATP synthesis, cortical granules (CGs) dynamic, F-actin polymerization, mitochondrial activity, mitochondrial damage, reactive oxygen species (ROS) production, intracellular glutathione (GSH) production, DNA damage, cellular apoptosis, fertilization capacity and blastocyst development potential of ovine oocytes. In addition, the oxidative stress damage model induced by HO treatment was applied to confirm the antioxidative effect of DMKG supplementation on the development of ovine oocytes. The results showed that compared with MII oocytes without DMKG supplementation (Control group), 3 mM DMKG supplementation during IVM significantly (P < 0.05) increased nuclear maturation rate, ATP synthesis, CGs dynamic, F-actin polymerization, mitochondrial activity, GSH production and embryonic developmental competence and decreased ROS production, mitochondrial damage, DNA damage and cellular apoptosis level of ovine MII oocytes. Moreover, the reductions in the developmental competences of ovine MII oocytes caused by HO induced oxidative stress damages were effectively ameliorated by the co-supplementation in IVM of 3 mM DMKG (P < 0.05). Our results demonstrate the promising effect of DMKG supplementation on the in vitro developmental competence of ovine oocytes via the reduction of oxidative stress damages and indicates further research into the clinical applications of DMKG and the development of ovine breeding technologies is warranted.
在体外成熟（IVM）过程中添加α-酮戊二酸二甲酯（DMKG）已被证明可提高猪卵母细胞的体外发育能力。在此，通过分析绵羊卵母细胞到中期II（MII）阶段的核成熟率、ATP合成、皮质颗粒（CGs）动态、F-肌动蛋白聚合、线粒体活性、线粒体损伤、活性氧（ROS）产生、细胞内谷胱甘肽（GSH）产生、DNA损伤、细胞凋亡、受精能力和囊胚发育潜力，研究了在IVM培养基中添加DMKG对绵羊卵母细胞发育能力的影响。此外，应用过氧化氢（HO）处理诱导的氧化应激损伤模型来证实添加DMKG对绵羊卵母细胞发育的抗氧化作用。结果表明，与未添加DMKG的MII卵母细胞（对照组）相比，在IVM期间添加3 mM DMKG显著（P < 0.05）提高了绵羊MII卵母细胞的核成熟率、ATP合成、CGs动态、F-肌动蛋白聚合、线粒体活性、GSH产生和胚胎发育能力，并降低了ROS产生、线粒体损伤、DNA损伤和细胞凋亡水平。此外，在IVM中共同添加3 mM DMKG可有效改善HO诱导的氧化应激损伤对绵羊MII卵母细胞发育能力的降低（P < 0.05）。我们的结果证明了添加DMKG通过减少氧化应激损伤对绵羊卵母细胞体外发育能力具有显著作用，并表明有必要进一步研究DMKG的临床应用和绵羊育种技术的发展。
