AAPH诱导的氧化应激对小鼠早期胚胎发育的作用及机制研究  

The Effect of Oxidative Stress Induced by AAPH on the Development of Mouse Preimplantation Embryos

作　　者：刘雅贤 于海泉 LIU Ya-xian;YU Hai-quan(The State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock,Inner Mongolia University,Hohhot 010070,China)

机构地区：[1]内蒙古大学省部共建草原家畜生殖调控与繁育国家重点实验室

出　　处：《内蒙古大学学报（自然科学版）》2019年第5期467-478,共12页Journal of Inner Mongolia University：Natural Science Edition

基　　金：国家自然科学基金项目(31760333,31460311)资助项目

摘　　要：哺乳动物胚胎发育是一个受复杂信号通路严格调控的过程,同时受遗传和环境因素等多种因素影响.由体外培养环境破坏活性氧ROS平衡所引起的氧化应激反应被认为是影响体外胚胎发育的重要因素之一,但活性氧影响胚胎的机制仍有待研究.本研究首先观察了叠氮化合物2,2’-偶氮二(2-脒基丙烷)盐酸盐AAPH[2,2-Azobis(2-Amidinopropane)dihydrochloride]处理后对胚胎发育的影响,检测了胚胎的活性氧水平、线粒体膜电位和细胞凋亡情况,同时检测了合子基因组激活相关基因的表达变化,针对活性氧诱导剂AAPH影响小鼠早期胚胎发育的分子机制进行研究.结果表明,AAPH能够提高小鼠胚胎的活性氧水平,影响胚胎发育率,AAPH的作用呈现剂量依赖性,这种影响是通过降低线粒体的膜电位,进而促发细胞凋亡.同时,抑制合子基因组激活相关基因Zscan4d、eIF-1A、Hspa1b、H2afz的表达.本研究探讨了活性氧影响早期胚胎发育过程的机制,为哺乳动物体外受精培养体系的优化提供了依据.Mammalian embryo development is a precisely regulated process that is governed by integrated signaling pathways and influenced by combinations of genetic and environmental factors.The oxidative stress reaction is considered an important factor which may destroy the balance of reactive oxygen species(ROS)in embryo.However,the molecular mechanism and the function of ROS remains unclear.In this study,the effect and the mechanism of oxidative stress,which induced by reactive oxygen revulsant AAPH,on mouse early embryos was evaluated.Firstly,the embryo development rates were observed after AAPH treatment at zygote stage,and then the concentration of ROS in embryo cytoplasm and the expression of related genes were detected.Secondly,the mitochondrial membrane potential and cell apoptosis were detected.Then the expression of ZGA related genes were examined.The results showed that a significant increase in the level of ROS in mouse embryo were observed after AAPH treatment.The development rate was affected in a dose-dependent way,the mitochondrial membrane potential of embryo decreased,and early apoptotic signal was detected.The expression levels of genes associated with zygotic genome activation was significantly decreased(P<0.01),including Zscan4 d,eIF-1 A,Hspa1 band H2 afz.This study preliminarily explored the molecular mechanism of ROS on mouse early embryonic development,which provided a reference for the optimization of mammalian embryo culture system.

关 键 词：活性氧 AAPH 合子基因组激活 胚胎发育 

分 类 号：Q952[生物学—动物学]