Department of Human Anatomy, Histology and Embryology, School of Basic Medical Sciences, Fujian Medical University, Fuzhou, Fujian 350122, China.
Anatomy Teaching Office, Basic Medical Science Department, Fujian Health College, Fuzhou, Fujian 350101, China.
Oxid Med Cell Longev. 2017;2017:2704532. doi: 10.1155/2017/2704532. Epub 2017 Jun 7.
During in vitro cultivation of preimplantation embryos, the balance between ROS production and clearance is disturbed and may lead to incompetent embryos, which might be a main reason of IVF-ET failure. Icariin (ICA) is reported to be active in clearing ROS. The present study aimed to investigate whether ICA could reverse HO pretreatment-induced mouse preimplantation embryo development arrest and, furthermore, to study the underlying mechanisms by detecting ROS levels, mitochondrial membrane potential (Δm), and zygotic gene expression. The results showed that, after pretreating mouse 1-cell embryos with 40 M or 60 M HO for 30 min, the developmental rate of each stage embryos decreased obviously. And by adding 40 M ICA, the developmental arrest of 60 M HO pretreated preimplantation embryos was significantly reversed. Immunostaining results showed that, comparing with the control group, ROS levels of HO pretreated 1-cell embryos were elevated and Δm levels decreased. By adding ICA, the ROS levels of HO pretreated 1-cell embryos were decreased and Δm levels were elevated. Furthermore, RT-qPCR results showed that the addition of ICA reversed the HO-induced downregulation of eIF-1A mRNA expression levels. These results indicate that ICA, when used in appropriate concentration, could decrease ROS levels, increase Δm levels, and modulate the expression of zygotic gene activation (ZGA) marker gene eIF-1A, and thus promote the development of HO-pretreated mouse preimplantation embryos.
在体外培养胚胎前，活性氧（ROS）的产生和清除之间的平衡被打破，可能导致胚胎发育能力不足，这可能是体外受精-胚胎移植（IVF-ET）失败的主要原因。淫羊藿苷（ICA）被报道具有清除 ROS 的作用。本研究旨在探讨 ICA 是否可以逆转 HO 预处理诱导的小鼠胚胎发育停滞，并通过检测 ROS 水平、线粒体膜电位（Δm）和合子基因表达来研究其潜在机制。结果表明，用 40μM 或 60μM HO 预处理小鼠 1 细胞胚胎 30min 后，各阶段胚胎的发育率明显下降。用 40μM ICA 处理，可明显逆转 60μM HO 预处理胚胎的发育停滞。免疫荧光染色结果显示，与对照组相比，HO 预处理的 1 细胞胚胎 ROS 水平升高，Δm 水平降低。加入 ICA 后，HO 预处理的 1 细胞胚胎 ROS 水平降低，Δm 水平升高。此外，RT-qPCR 结果表明，ICA 的加入逆转了 HO 诱导的 eIF-1A mRNA 表达水平下调。这些结果表明，ICA 在适当浓度下可降低 ROS 水平，增加Δm 水平，并调节合子基因激活（ZGA）标记基因 eIF-1A 的表达，从而促进 HO 预处理小鼠胚胎的发育。
