Lee Sanghoon, Park Eun Jung, Moon Joon Ho, Kim Su Jin, Song Kilyoung, Lee Byeong Chun
Department of Theriogenology and Biotechnology, Research Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University, Seoul, Republic of Korea.
Department of Theriogenology and Biotechnology, Research Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University, Seoul, Republic of Korea; Institute of Green Bio Science and Technology, Seoul National University, Pyeongchang, Gangwon-do, Korea.
Theriogenology. 2015 Jul 1;84(1):145-54. doi: 10.1016/j.theriogenology.2015.02.021. Epub 2015 Mar 5.
We investigated the effect of resveratrol supplementation during IVM and/or trolox during IVC on the development of porcine embryos derived from parthenogenetic activation (PA) and SCNT. In this study, we evaluated intracellular glutathione (GSH) and reactive oxygen species (ROS) levels, gene expression in blastocysts, and embryonic development after PA and SCNT. To determine the combined effects of resveratrol during IVM and trolox during IVC on PA embryos, we selected optimal concentrations (2 μM of resveratrol and 200 μM of trolox) and designed four groups: (1) control, (2) resveratrol, (3) trolox, and (4) combined. All treatment groups showed significantly increased intracellular GSH levels and decreased ROS levels. Resveratrol supported significantly higher cleavage and blastocyst formation rates than the control (80.3% and 38.0% vs. 71.1% and 22.4%, respectively) by downregulating Bax/Bcl-2, Caspase-3, and Bak. Trolox showed significantly increased blastocyst formation rates (36.7%) compared with the control (22.4%) by downregulating only Caspase-3. The combined group had significantly higher cleavage and blastocyst formation rates and greater total cell numbers than the control (81.7%, 36.3%, and 67.1 vs. 71.1%, 22.4%, and 47.8, respectively) by downregulating Bax/Bcl-2, Caspase-3, and Bak. On the basis of these results, we applied sequential treatments with resveratrol and trolox to SCNT, and blastocyst formation rates and total cell numbers were significantly increased compared with the control (17.2% and 52.1 vs. 11.8% and 36.6, respectively), with increased GSH, decreased ROS levels, upregulated proliferating cell nuclear antigen, and downregulated Bax/Bcl-2 and Caspase-3. These results indicate that sequential treatment with resveratrol during IVM and trolox during IVC improved the development of PA and SCNT porcine embryos by regulating intracellular GSH, ROS levels, and gene expression.
我们研究了体外成熟（IVM）期间补充白藜芦醇和/或体外培养（IVC）期间补充生育三烯酚对孤雌激活（PA）和体细胞核移植（SCNT）来源的猪胚胎发育的影响。在本研究中，我们评估了细胞内谷胱甘肽（GSH）和活性氧（ROS）水平、囊胚中的基因表达以及PA和SCNT后的胚胎发育。为了确定IVM期间白藜芦醇和IVC期间生育三烯酚对PA胚胎的联合作用，我们选择了最佳浓度（2 μM白藜芦醇和200 μM生育三烯酚）并设计了四组：（1）对照组，（2）白藜芦醇组，（3）生育三烯酚组，和（4）联合组。所有处理组的细胞内GSH水平均显著升高，ROS水平均降低。白藜芦醇通过下调Bax/Bcl-2、Caspase-3和Bak，支持的卵裂率和囊胚形成率显著高于对照组（分别为80.3%和38.0%，而对照组为71.1%和22.4%）。生育三烯酚仅通过下调Caspase-3，与对照组（22.4%）相比，囊胚形成率显著提高（36.7%）。联合组通过下调Bax/Bcl-2、Caspase-3和Bak，其卵裂率和囊胚形成率显著高于对照组，总细胞数也更多（分别为81.7%、36.3%和67.1，而对照组为71.1%、22.4%和47.8）。基于这些结果，我们将白藜芦醇和生育三烯酚的序贯处理应用于SCNT，与对照组相比，囊胚形成率和总细胞数显著增加（分别为17.2%和52.1，而对照组为11.8%和36.6），同时GSH增加，ROS水平降低，增殖细胞核抗原上调，Bax/Bcl-2和Caspase-3下调。这些结果表明，IVM期间白藜芦醇和IVC期间生育三烯酚的序贯处理通过调节细胞内GSH、ROS水平和基因表达，改善了PA和SCNT猪胚胎的发育。
