Protective Effect of Icariin on the Development of Preimplantation Mouse Embryos against Hydrogen Peroxide-Induced Oxidative Injury.

Rong Ye,Yang Su,Yuhuan Zhong,Xiuli Lian,Songhua Xu,Shie Wang,Lili Pang,Yue Liu

doi:10.1155/2017/2704532

Abstract

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 H2O2 pretreatment-induced mouse preimplantation embryo development arrest and, furthermore, to study the underlying mechanisms by detecting ROS levels, mitochondrial membrane potential (ΔΨm), and zygotic gene expression. The results showed that, after pretreating mouse 1-cell embryos with 40 μM or 60 μM H2O2 for 30 min, the developmental rate of each stage embryos decreased obviously. And by adding 40 μM ICA, the developmental arrest of 60 μM H2O2 pretreated preimplantation embryos was significantly reversed. Immunostaining results showed that, comparing with the control group, ROS levels of H2O2 pretreated 1-cell embryos were elevated and ΔΨm levels decreased. By adding ICA, the ROS levels of H2O2 pretreated 1-cell embryos were decreased and ΔΨm levels were elevated. Furthermore, RT-qPCR results showed that the addition of ICA reversed the H2O2-induced downregulation of eIF-1A mRNA expression levels. These results indicate that ICA, when used in appropriate concentration, could decrease ROS levels, increase ΔΨm levels, and modulate the expression of zygotic gene activation (ZGA) marker gene eIF-1A, and thus promote the development of H2O2-pretreated mouse preimplantation embryos.

Highlights

After about thirty years of development, human-assisted reproduction technology, such as in vitro fertilization and embryo transplantation (IVF-ET), has gained great progress
Mouse 1-cell embryos were collected, cultured in 60 μM H2O2 for 30 min, washed for 3 times, and moved into KSOM medium supplemented with different final concentrations of ICA (0 μM, 10 μM, 20 μM, 40 μM, and 80 μM) for further cultivation until blastocyst stage
To investigate the reactive oxygen species (ROS) clearing functions of ICA in preimplantation embryos, mouse 1-cell embryos were pretreated with 60 μM H2O2 for 30 min and moved into KSOM medium supplemented with different final concentrations of ICA

Summary

Introduction

After about thirty years of development, human-assisted reproduction technology, such as in vitro fertilization and embryo transplantation (IVF-ET), has gained great progress. The lacking of competent early embryos for transplantation usually forces the infertile couples to abandon IVF-ET. It is of need to improve the in vitro cultivation system, to help the “weak” preimplantation embryos overcome developmental arrest. Of the various harmful factors that may result in developmental blockage, the effects induced by excessive reactive oxygen species (ROS) are considered to be among the most significant ones [1, 2]. While, during the consumption of oxygen, derivatives of oxygen that have active chemical reactivity (and be termed as ROS) are produced, ((inNHcO2lOu−d2)i).n, gWhhysdeunrpoextrhyolexidforeveeeraprnraoidodniucacle(dO(O2R−H)O,−S)h,byradenraodkgentnhitrroipucegrhooxxtiihddeee defensive walls of the organism, it will result in oxidative

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Conclusion