Addition of glutathione or thioredoxin to culture medium reduces intracellular redox status of porcine IVM/IVF embryos, resulting in improved development to the blastocyst stage
Addition of glutathione or thioredoxin to culture medium reduces intracellular redox status of porcine IVM/IVF embryos, resulting in improved development to the blastocyst stage
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DOI：
10.1002/mrd.20533
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发表时间：
2006-08-01
影响因子：
2.5
通讯作者：
Kikuchi, Kazuhiro
中科院分区：
文献类型：
--
作者：
Ozawa, Manabu;Nagai, Takashi;Kikuchi, Kazuhiro
本系列实验研究了通过在体外培养液中添加还原型谷胱甘肽（GSH）或硫氧还蛋白（TRX）所产生的还原环境对体外生产的猪胚胎发育能力及其细胞内氧化还原状态的影响。从体外成熟和受精的猪卵巢中收集卵丘-卵母细胞复合体。然后将推定的合子在含有或不含有（对照）GSH或TRX的改良NCSU-37培养基中培养6天，并评估其发育能力。此外，使用基于细胞内GSH含量相对于细胞内H2 O2水平的比率的指数定量地比较培养胚胎的细胞内氧化还原状态。在培养液中添加0.5或1.0 μ M GSH（29.6%或30.4%，P < 0.05或0.01）或1.0 mg/ml TRX（30.6%，P < 0.01）时，发育到囊胚期的胚胎比例显著高于不添加任何GSH的对照组（20.1%）。与对照组相比，在GSH或TRX存在下培养的8- 12-细胞期或囊胚期胚胎的细胞内氧化还原状态显著降低（P < 0.05至0.001）。与对照组相比，GSH或TRX处理组囊胚细胞总数增加（从48.3增加到49.2），凋亡细胞比例降低（从6.2%降低到7.0%）（细胞总数为39.3，凋亡率为11.1%，P < 0.05）。这些结果表明，GSH或TRX可以促进猪胚胎的体外发育，同时保持细胞内还原状态。
The present series of experiments investigated the effect of a reducing environment created by addition of reduced glutathione (GSH) or thioredoxin (TRX) to in vitro culture medium on the developmental competence of in vitro produced porcine embryos, and their intracellular redox status. Porcine cumulus-oocyte complexes were collected from ovaries matured and fertilized in vitro. The putative zygotes were then cultured for 6 days in modified NCSU-37 medium with or without (control) GSH or TRX, and their developmental competence was evaluated. In addition, the intracellular redox status of the cultured embryos was compared quantitatively using an index based on the ratio of the intracellular GSH content relative to the intracellular H2O2 level. The proportion of embryos that developed to the blastocyst stage was significantly increased when 0.5 or 1.0 mu M GSH (29.6% or 30.4%, P < 0.05 or 0.01, respectively) or 1.0 mg/ml TRX (30.6%, P < 0.01) was added to the medium compared to that without any supplementation (control; 20.1%). The intracellular redox status of embryos at the 8- to 12-cell stage or the blastocyst stage in the group cultured in the presence of GSH or TRX was significantly reduced in comparison with the control (P < 0.05 to 0.001). Furthermore, administration of GSH or TRX enhanced the total cell number (from 48.3 to 49.2) and lowered the proportion of apoptotic cells (from 6.2% to 7.0%) in blastocysts compared with the control (cell number 39.3; apoptosis rate 11.1%, P < 0.05). These results suggest that GSH or TRX can improve the in vitro development of porcine embryos, while maintaining an intracellular reductive status.