白藜芦醇对猪卵母细胞体外成熟以及孤雌激活和体外受精后胚胎发育的影响。
The effects of resveratrol on porcine oocyte in vitro maturation and subsequent embryonic development after parthenogenetic activation and in vitro fertilization.
机构信息
Laboratory of Veterinary Embryology and Biotechnology, College of Veterinary Medicine, Chungbuk National University, Chungbuk, South Korea.
出版信息
Theriogenology. 2012 Jul 1;78(1):86-101. doi: 10.1016/j.theriogenology.2012.01.024. Epub 2012 Mar 23.
We investigated the effects of resveratrol, a phytoalexin with various pharmacologic activities, on in vitro maturation (IVM) of porcine oocytes. We investigated intracellular glutathione (GSH) and reactive oxygen species (ROS) levels, as well as gene expression in mature oocytes, cumulus cells, and in vitro fertilization (IVF)-derived blastocysts, and subsequent embryonic development after parthenogenetic activation (PA) and IVF. After 44 h of IVM, no significant difference was observed in maturation of the 0.1, 0.5, and 2.0 μM resveratrol groups (83.0%, 84.1%, and 88.3%, respectively) compared with the control (84.1%), but the 10.0 μM resveratrol group showed significantly decreased nuclear maturation (75.0%) (P < 0.05). The 0.5- and 2.0-μm groups showed a significant (P < 0.05) increase in intracellular GSH levels compared with the control and 10.0 μM group. Intracellular ROS levels in oocytes matured with 2.0 μM resveratrol decreased significantly (P < 0.05) compared with those in the other groups. Oocytes treated with 2.0 μM resveratrol during IVM had significantly higher blastocyst formation rates and total cell numbers after PA (62.1% and 49.1 vs. 48.8%, and 41.4, respectively) and IVF (20.5% and 54.0 vs. 11.0% and 43.4, respectively) than the control group. Cumulus-oocytes complex treated with 2.0 μM resveratrol showed lower expression of apoptosis-related genes compared with mature oocytes and cumulus cells. Cumulus cells treated with 2.0 μM resveratrol showed higher (P < 0.05) expression of proliferating cell nuclear antigen than the control group. IVF-derived blastocysts derived from 2.0 μM resveratrol-treated oocytes also had less (P < 0.05) Bak expression than control IVF-derived blastocysts. In conclusion, 2.0 μM resveratrol supplementation during IVM improved the developmental potential of PA and IVF porcine embryos by increasing the intracellular GSH level, decreasing ROS level, and regulating gene expression during oocyte maturation.
我们研究了白藜芦醇对猪卵母细胞体外成熟（IVM）的影响。我们研究了成熟卵母细胞、卵丘细胞以及体外受精（IVF）衍生的囊胚中的细胞内谷胱甘肽（GSH）和活性氧（ROS）水平以及基因表达情况，随后对孤雌激活（PA）和 IVF 后的胚胎发育情况进行了研究。在 IVM 44 小时后，与对照组（84.1%）相比，0.1、0.5 和 2.0μM 白藜芦醇组的成熟率（分别为 83.0%、84.1%和 88.3%）无显著差异，但 10.0μM 白藜芦醇组的核成熟率显著降低（75.0%）（P<0.05）。与对照组和 10.0μM 组相比，0.5-和 2.0-μm 组的细胞内 GSH 水平显著升高（P<0.05）。与其他组相比，用 2.0μM 白藜芦醇成熟的卵母细胞中的细胞内 ROS 水平显著降低（P<0.05）。IVM 期间用 2.0μM 白藜芦醇处理的卵母细胞在 PA（62.1%和 49.1%，分别为 48.8%和 41.4%）和 IVF（20.5%和 54.0%，分别为 11.0%和 43.4%）后囊胚形成率和总细胞数均显著升高（P<0.05）。与成熟卵母细胞和卵丘细胞相比，用 2.0μM 白藜芦醇处理的卵丘-卵母细胞复合物中凋亡相关基因的表达水平较低。与对照组相比，用 2.0μM 白藜芦醇处理的卵丘细胞中增殖细胞核抗原的表达水平更高（P<0.05）。用 2.0μM 白藜芦醇处理的 IVF 衍生囊胚中 Bak 基因的表达水平也低于对照组的 IVF 衍生囊胚（P<0.05）。总之，在 IVM 期间补充 2.0μM 白藜芦醇可通过增加细胞内 GSH 水平、降低 ROS 水平以及调节卵母细胞成熟过程中的基因表达来提高 PA 和 IVF 猪胚胎的发育潜力。
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