芝麻素对小鼠早期胚胎体外发育的影响

Effect of Sesamin on the Development of Early Mice Embryo in vitro

本试验旨研究在体外培养(in vitro culture,IVC)中添加芝麻素(sesamin,SES)对小鼠早期胚胎体外发育的影响。采集6周龄雌性昆明小鼠受精卵,随机分为对照组和不同浓度(10、50、100μmol/L)SES组。采用Fluorescein-dUTP和Hoechst 33342免疫荧光染色分别检测囊胚内细胞凋亡率和总细胞数;采用DCFH-DA检测早期胚胎内活性氧(reactive oxygen species,ROS)水平;采用CMF 2HC检测早期胚胎内谷胱甘肽(glutathione,GSH)水平;采用JC-1检测早期胚胎线粒体膜电位强度。结果显示,不同浓度SES组分裂率和囊胚率较对照组均有提高,但无显著差异(P>0.05);50μmol/L SES组细胞数较对照组显著提高(P<0.05),细胞凋亡率较对照组显著降低(P<0.05);与对照组相比,50μmol/L SES组ROS水平显著降低(P<0.05),GSH水平和线粒体膜电位水平显著提高(P<0.05)。结果表明,在IVC中添加SES可提高囊胚内细胞数、减少细胞凋亡率,降低细胞内ROS水平,提高细胞内GSH水平,改善早期胚胎线粒体功能,减少胚胎氧化应激的损伤,提高小鼠早期胚胎发育质量。

The aim of this study was to investigate the effect of sesamin(SES)added to in vitro culture(IVC)on the development of early mice embryos.The fertilized eggs of 6-week-old female Kunming mice were randomly divided into control group and different concentrations(10,50,100μmol/L)SES groups.The apoptosis rate and total cell number in blastocysts were detected by Fluorescein-dUTP and Hoechst 33342 immunofluorescence staining,respectively.DCFH-DA was used to detect the levels of reactive oxygen species(ROS)in early embryos;The levels of glutathione(GSH)in early embryos were detected by CMF 2HC;Early embryonic mitochondrial membrane potential intensity was measured using JC-1.The results showed that the segregation rate and blastocyst rate of SES groups were higher than those in control group,but there was no significant difference(P>0.05).The number of cells in 50μmol/L SES group was significantly higher than that in control group(P<0.05),and the apoptosis rate was significantly lower than that in control group(P<0.05).Compared with control group,the ROS level in the 50μmol/L SES group was significantly lower(P<0.05),and the GSH level and mitochondrial membrane potential level were significantly increased(P<0.05).The results showed that the addition of SES in IVC could increase the number of cells in blastocyst,decrease the apoptotic rate and the ROS level,increase the GSH level,improve the mitochondrial function of early embryos,reduce the damage of oxidative stress of embryos,and improve the quality of early embryonic development in mice.