人类胚胎线粒体ATP、ROS水平与胚胎形态学分型的关系

Relationship between Levels of ATP and ROS of Human Embryo Mitochondria and Embryo Morphology

目的探讨人类胚胎线粒体三磷酸腺苷(ATP)、胚胎活性氧(ROS)水平与第3天(D3)不同形态学分型胚胎的关系及其对胚胎发育的影响。方法选择行体外受精-胚胎移植(IVF-ET)的不孕患者新鲜周期发育较差不适合移植的胚胎和助孕成功妊娠后自愿丢弃的质量较好的2PN冷冻胚胎。根据国际胚胎评分共识标准,将卵裂期胚胎分为Ⅰ级冷冻胚胎组、Ⅱ级冷冻胚胎组和Ⅲ级新鲜胚胎组(不能移植的胚胎)。采用多功能酶标仪和共聚焦显微镜分别检测单个胚胎ATP和ROS表达水平,比较D3不同形态学分型的胚胎ATP水平和ROS浓度的差异。结果Ⅲ级新鲜胚胎组ATP水平明显低于Ⅰ级冷冻胚胎组和Ⅱ级冷冻胚胎组(均P<0.05),ROS浓度明显高于Ⅰ级冷冻胚胎组和Ⅱ级冷冻胚胎组(均P<0.05),Ⅰ级冷冻胚胎组ATP水平明显高于Ⅱ级冷冻胚胎组(P<0.05),ROS浓度明显低于Ⅱ级冷冻胚胎组(P<0.05)。结论质量越好的D3胚胎ATP水平越高,ROS水平越低。

Objective To investigate the relationship between the embryos with different morphological types on day 3（D3）and embryo mitochodria ATP and ROS,and their influence on embryonic development.Methods We selected embryos with poor development after normal fertilization and not suitable for transplant in fresh cycle and frozen embryos were discarded voluntarily by infertility patients with IVF-ET after successful pregnancy.According to international standard of embryo evaluation,all cleavage stage embryos were divided into classⅠfrozen embryo group,classⅡfrozen embryo group and classⅢfresh embryo group（embryos unable to be transplanted）.And then the luciferase assay was used to detect single embryo ATP levels,activated oxygen detection kit to detect the ROS level of a single embryo.The differences of ATP level and ROS concentration were compared between the embryos with different morphological types on D3.Results The mitochondrial ATP of class Ⅲfresh embryonic group was significantly lower than that of class Ⅰ and Ⅱ frozen embryo groups（both P〈0.05）.The mitochondrial ROS of classⅢ fresh embryonic group was significantly higher than that of class Ⅰ and Ⅱ frozen embryo groups（both P〈0.05）.The mitochondrial ATP of classⅠfrozen embryo group was significantly higher than that of classⅡ frozen embryo group（P〈0.05）.The mitochondrial ROS of classⅠfrozen embryo group was significantly lower than that of classⅡfrozen embryo group（P〈0.05）.Conclusion The higher the quality of embryos on D3,the higher the ATP and the lower the ROS level in embryos.