Melatonin inhibits apoptosis and improves the developmental potential of vitrified bovine oocytes
Melatonin inhibits apoptosis and improves the developmental potential of vitrified bovine oocytes
复制标题
褪黑素抑制细胞凋亡并提高玻璃化牛卵母细胞的发育潜力
DOI：
10.1111/jpi.12290
复制
发表时间：
2016-03-01
影响因子：
10.3
通讯作者：
Zhu, Hua-Bin
中科院分区：
文献类型：
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作者：
Zhao, Xue-Ming;Hao, Hai-Sheng;Zhu, Hua-Bin
卵母细胞的玻璃化冷冻已被证明与活性氧(ROS)水平的升高和细胞凋亡事件密切相关。然而，关于褪黑素对玻璃化冷冻卵母细胞中ROS水平和凋亡事件的影响的信息很少。因此，我们通过添加玻璃化冷冻液或体外成熟液以及玻璃化冷冻液中添加10−-9M褪黑素来研究褪黑素对牛卵母细胞玻璃化冷冻后ROS和细胞凋亡的影响。分析了牛卵母细胞玻璃化冷冻后的ROS、线粒体钙离子(mCa~(2+))和膜电位(ΔΨ_m)、磷脂酰丝氨酸(PS)外在化、半胱氨酸天冬氨酸氨基转移酶-3(Caspase-3)活化、DNA片段化、Bax和Bcl2L1mRNA表达水平以及发育潜能。玻璃化冷冻后的牛卵母细胞ROS、mCa~(2+)、Bax mRNA和Caspase-3蛋白水平升高，PS外化率和DNA片段率升高，ΔΨm和Bcl2 L1mRNA表达水平降低。在玻璃化液中添加褪黑激素后，卵母细胞的ROS、mCa~(2+)、Bax基因表达、caspase-3蛋白水平、PS外化率和DNA碎片率显著降低，ΔΨm和Bcl2 L1基因表达水平显著升高，从而提高了玻璃化冷冻卵母细胞孤雌激活后的发育能力。添加褪黑激素的玻璃化冷冻卵母细胞在体外成熟液和玻璃化冷冻液中的发育能力与新鲜卵母细胞相似。本研究表明，添加10−-9M褪黑素的牛卵母细胞玻璃化冷冻液或玻璃化冷冻液可显著降低玻璃化冷冻牛卵母细胞的ROS水平，抑制其细胞凋亡，从而提高其发育潜力。
Vitrification of oocytes has been shown to be closely associated with increased levels of reactive oxygen species (ROS) and apoptotic events. However, little information is available the effect of melatonin on the ROS levels and apoptotic events in vitrified oocytes. Therefore, we studied the effect of melatonin on ROS and apoptotic events in vitrified bovine oocytes by supplementing vitrification solution or in vitro maturation (IVM) and vitrification solution with 10−9 m melatonin. We analyzed the ROS, mitochondrial Ca2+ (mCa2+) and membrane potential (ΔΨm), externalization of phosphatidylserine (PS), caspase‐3 activation, DNA fragmentation, mRNA expression levels of Bax and Bcl2 l1, and developmental potential of vitrified bovine oocytes. Vitrified bovine oocytes exhibited increased levels of ROS, mCa2+, Bax mRNA, and caspase‐3 protein and higher rates of PS externalization and DNA fragmentation, and decreased ΔΨm and Bcl2 l1 mRNA expression level. However, melatonin supplementation in vitrification solution or IVM and vitrification solution significantly decreased the levels of ROS, mCa2+, Bax mRNA expression, and caspase‐3 protein, and PS externalization and DNA fragmentation rates, and increased the ΔΨm and Bcl2 l1 mRNA expression level in vitrified oocytes, resulting in an increased developmental ability of vitrified bovine oocytes after parthenogenetic activation. The developmental ability of vitrified oocytes with melatonin supplementation in IVM and vitrification solution was similar to that of fresh ones. This study showed that supplementing the IVM and vitrification medium or vitrification medium with 10−9 m melatonin significantly decreased the ROS level and inhibited apoptotic events of vitrified bovine oocytes, consequently increasing their developmental potential.