Effects of purple sweet potato anthocyanins on development and intracellular redox status of bovine preimplantation embryos exposed to heat shock
Effects of purple sweet potato anthocyanins on development and intracellular redox status of bovine preimplantation embryos exposed to heat shock
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DOI：
10.1262/jrd.18124
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发表时间：
2007-06-01
影响因子：
1.8
通讯作者：
Takahashi, Masashi
中科院分区：
文献类型：
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作者：
Sakatani, Miki;Suda, Ikuo;Takahashi, Masashi
卵裂期植入前胚胎的发育因暴露于热休克而中断，例如夏季的高温。在这项研究中，我们研究了是否添加花青素，这是活性氧（ROS）的强清除剂，改善热暴露的牛植入前胚胎的发育和细胞内氧化还原状态，通过减少热休克衍生的氧化应激。在体外受精（IVF）后，胚胎在38.5 ℃下用0、0.1、1和10 μ g/ml花青素（非热休克组）培养至第8天（第0天= IVF当天）。在第2天，将胚胎在41.5 ℃下用0、0.1、1和10 μ g/ml花青素苷培养6小时，然后在38.5 ℃下培养直到第8天（HS组）。热休克处理后，用荧光探针测定非热休克组和热休克组胚胎细胞内活性氧和谷胱甘肽（GSH）含量。在第8天，评价胚胎的囊胚形成率和囊胚的总细胞数。与非热休克胚胎相比，不含花青素的热休克胚胎的囊胚形成率和GSH含量显著降低（P <0.05），细胞内ROS含量显著升高（P < 0.05）。相反，添加0.1 μ g/ml花青素显著提高了热休克胚胎的囊胚形成率（P < 0.05）。在热激条件下，添加任何剂量的花青素均能显著降低ROS水平（P < 0.05），并有升高GSH水平的趋势。然而，除了较高浓度（1和10 μ g/ml）的花青素的培养基下热休克没有改善胚胎的发育。这些结果表明，花青素通过降低细胞内氧化应激和增加GSH水平维持热休克牛胚胎的细胞内氧化还原平衡。因此，使用天然抗氧化多酚改变氧化还原状态是减少热休克衍生的氧化应激的有用方法。
The development of cleavage stage preimplantation embryos is disrupted by exposure to heat shock, such as high temperatures in the summer season. In this study, we investigated whether addition of anthocyanins, which are strong scavengers of reactive oxygen species (ROS), improves development and intracellular redox status of heat-exposed bovine preimplantation embryos by reduction of heat shock-derived oxidative stress. After in vitro fertilization (IVF), embryos were cultured at 38.5 C through Day 8 (Day 0 = day of IVF) with 0, 0.1, 1 and 10 mu g/ml anthocyanins (non-heat-shocked group). On Day 2, embryos were cultured at 41.5 C for 6 h with 0, 0.1, 1 and 10 mu g/ml anthocyanins followed by culture at 38.5 C until Day 8 (HS group). After exposure to heat shock, the intracellular ROS and glutathione (GSH) contents of individual embryos were measured in the non-heat-shocked and HS groups using fluorescent probes. On Day 8, the blastocysts formation rates of the embryos and total cell numbers of blastocysts were evaluated. Embryos exposed to heat shock without anthocyanins showed a significant decrease in blastocyst formation rate and GSH content (P < 0.05) and an increase in intracellular ROS (P < 0.05) compared with non-heat-shocked embryos. In contrast, addition of 0.1 mu g/ml anthocyanins significantly (P < 0.05) improved the blastocyst formation rate of the heat-shocked embryos. Addition of any dose of anthocyanins produced a significant decrease in the ROS levels (P < 0.05) and tended to increase the GSH levels under heat-shock conditions. However, addition of higher concentrations (1 and 10 mu g/ml) of anthocyanins to the culture media under heat shock did not improve the development of embryos. These results indicate that anthocyanins maintain the intracellular redox balance of heat-shocked bovine embryos by reducing intracellular oxidative stress and increasing the GSH levels. Thus, alterations of the redox state using natural antioxidative polyphenols is a useful approach for reducing heat shock-derived oxidative stress.