Glutathione is present in reproductive tract secretions and improves development of mouse embryos after chemically induced glutathione depletion
Glutathione is present in reproductive tract secretions and improves development of mouse embryos after chemically induced glutathione depletion
复制标题
DOI：
10.1095/biolreprod59.2.431
复制
发表时间：
1998-08-01
影响因子：
3.6
通讯作者：
Stover, SK
中科院分区：
文献类型：
--
作者：
Gardiner, CS;Salmen, JJ;Stover, SK
我们调查的假设，即还原型谷胱甘肽（GSH）是存在于女性生殖道的分泌物，这种细胞外GSH可能会保护preimprantation小鼠胚胎细胞内GSH耗尽后。卵裂期小鼠胚胎不能重新合成谷胱甘肽，也不能从体外谷胱甘肽耗竭中恢复。通过HPLC和Bradford方法分别定量分析输卵管冲洗液中的GSH和总蛋白，发现每毫克总蛋白中有51 nmol GSH。用60 μ M马来酸二乙酯（DEM）处理胚胎以耗尽细胞GSH。当用ImM GSH培养时，与在对照培养基中培养的那些相比，这些胚胎表现出改善的发育（96%对87%桑椹胚[p < 0.05]，78%对75%囊胚，58%对54%扩张囊胚，21%对17%起始孵化囊胚）。然而，细胞内的GSH含量的胚胎没有显着增加的DEM处理的胚胎在培养基中含有GSH的培养16，40，或64小时，这表明胚胎是不能采取完整的GSH。此外，向培养基中加入丁硫丙啶亚砜亚胺（抑制GSH合成）或阿西霉素（抑制GSH在膜上分解）阻断了发育的改善。这些数据表明，生殖道液中的GSH可能有助于保护植入前胚胎免受毒性和内源性胚胎GSH耗竭的不利影响。
We investigated the hypothesis that reduced glutathione (GSH) is present in secretions of the female reproductive tract and that this extracellular GSH may protect preimprantation mouse embryos after intracellular GSH depletion. The cleavage-stage mouse embryo cannot synthesize GSH de novo and is unable to recover from glutathione depletion in vitro. Analysis of GSH and total protein of oviduct flushings, quantified by HPLC and the Bradford method, respectively, revealed 51 nmol GSH per mg total protein. Embryos were treated with 60 mu M diethyl maleate (DEM) to deplete cellular GSH. When cultured with 1 mM GSH, these embryos exhibited improved development compared to those cultured in control medium (96% vs. 87% morula [p < 0.05], 78% vs. 75% blastocyst, 58% vs. 54% expanded blastocyst, 21% vs. 17% initiating hatching blastocyst). However, intracellular GSH content of embryos was not significantly increased by the culture of DEM-treated embryos in medium containing GSH for 16, 40, or 64 h of incubation, suggesting that the embryo is not capable of taking up intact GSH. Furthermore, addition of buthionine sulfoximine (which inhibits synthesis of GSH) or acivicin (which inhibits breakdown of GSH at the membrane) to culture medium blocked the improvement in development. These data suggest that GSH in reproductive tract fluid may help protect preimplantation embryos from the adverse effects of toxicant-induced and endogenous depletion of embryonic GSH.