Sequential treatment with resveratrol-trolox improves development of porcine embryos derived from parthenogenetic activation and somatic cell nuclear transfer
Sequential treatment with resveratrol-trolox improves development of porcine embryos derived from parthenogenetic activation and somatic cell nuclear transfer
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DOI：
10.1016/j.theriogenology.2015.02.021
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发表时间：
2015-07-01
期刊：
影响因子：
2.8
通讯作者：
Lee, Byeong Chun
中科院分区：
文献类型：
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作者：
Lee, Sanghoon;Park, Eun Jung;Lee, Byeong Chun
我们研究了体外受精期间补充白藜芦醇和/或体外受精期间补充trolox对孤雌生殖激活（PA）和SCNT衍生的猪胚胎发育的影响。在这项研究中，我们评估了PA和SCNT后细胞内谷胱甘肽（GSH）和活性氧（ROS）水平、囊胚基因表达和胚胎发育。为了确定体外受精期间白藜芦醇和体外受精期间trolox对PA胚胎的联合影响，我们选择了最佳浓度（2 μ M白藜芦醇和200 μ M trolox），设计了4个组：(1)对照，(2)白藜芦醇，(3)trolox和(4)联合。所有治疗组细胞内GSH水平均显著升高，ROS水平显著降低。白藜芦醇通过下调Bax/Bcl-2、Caspase-3和Bak，使卵裂率和囊胚形成率显著高于对照组（分别为80.3%和38.0%，71.1%和22.4%）。与对照组（22.4%）相比，Trolox仅下调Caspase-3可显著提高囊胚形成率（36.7%）。联合组通过下调Bax/Bcl-2、Caspase-3和Bak，其卵裂率、囊胚形成率和总细胞数均显著高于对照组（分别为81.7%、36.3%和67.1比71.1%、22.4%和47.8）。在此基础上，我们对SCNT进行白藜芦醇和trolox序次处理，囊胚形成率和总细胞数较对照组显著增加（分别为17.2%和52.1比11.8%和36.6%），GSH升高，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，在体外受精期间依次使用白藜芦醇和体外受精期间依次使用trolox通过调节细胞内GSH、ROS水平和基因表达来促进PA和SCNT猪胚胎的发育。(C) 2015爱思唯尔公司版权所有。
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 mu M of resveratrol and 200 mu M 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. (C) 2015 Elsevier Inc. All rights reserved.