【作者】 何岩； 于洋； 董婉维； 周生来； 王惟； 杨葳； 郑志红；

【Author】 HE Yan;YU Yang;ZHOU Shenglai;WANG Wei;ZHENG Zhihong;Department of Laboratory Animal Science,China Medical University;

【机构】 中国医科大学实验动物部；

【摘要】 目的研究脱氢表雄酮(DHEA)对3周龄C57BL/6J雌鼠超排卵效果和卵母细胞质量影响及其机制。方法 3周龄C57BL/6J雌鼠30只随机分为3组,分别为对照组、低剂量处理组(2mg/kg·d-1)和高剂量处理组(5mg/kg·d-1)。用DHEA按照低剂量和高剂量给予低剂量处理组和高剂量处理组灌胃1周,再经激素刺激超排卵,收集卵母细胞分别进行体外受精和收集卵母细胞周围的卵丘细胞。提取收集的卵丘细胞RNA进行qRT-PCR检测与胚胎质量相关基因Grem1、Has2、Ptgs1、Ptgs2和Vcan的m RNA水平以及凋亡相关基因Caspase3和Bcl2l10的m RNA水平。结果经过DHEA处理的雌鼠超排卵后,体外受精(IVF)显示实验组异形卵比例下降,经过DHEA高剂量处理组比低剂量处理组异形卵比例更低。卵丘细胞中mRNA表达水平显示,DHEA处理组Grem1表达水平与对照组相比降低,且高剂量组比低剂量组降低的倍数更大。DHEA对于Ptgs2的m RNA水平与其对Grem1的表达水平有相似的结果,即实验组mRNA水平比对照组降低。而另外的胚胎质量相关标记物Has2、Vcan和Ptgs1的m RNA水平实验组与对照组比较显著上调。Has2上调最明显。差异均有统计学意义。卵丘细胞中凋亡相关基因Caspase3表达水平显示,DHEA处理组与实验组相比显著下调,高剂量组下调倍数比低剂量组更明显,即低剂量组下调0.8倍,高剂量组下调0.3倍。Bcl2l10的m RNA表达水平显示,实验组m RNA表达水平高于对照组。差异均有统计学意义。结论经过DHEA处理后可以通过抑制卵丘细胞凋亡,提高卵母细胞的质量,降低3周龄C57BL/6J雌鼠超排卵异形卵的比率,优化超排卵方法。更多还原

【Abstract】 Objective To investigate the effect and mechanism of DHEA on the quality of oocytes of 3 weeks C57 BL/6 J mice after superovulation Methods Thirty 3-week-old C57 BL/6 J female mice were randomly divided into three groups:control group,low-dose treatment group(2 mg/kg.d-1) and high-dose treatment group(5 mg/kg.d-1).DHEA was administrated via the gastrointestinal tract for one week.Oocytes were collected after superovulation and underwent IVF.qRT-PCR was used to detect gene expression of Grem1、Has2、Ptgs1、Ptgs2 and Vcan,the makers of cumulus cells collected after superovulation.Gene expression of Caspase3 and Bcl2 l10 were also detected by qRT-PCR.Results After superovulation of female mice treated with DHEA,in vitro fertilization(IVF) showed that the proportion of heteromorphic eggs in the experimental group decreased,and the proportion of heteromorphic eggs in the high-dose DHEA group was lower than that in the low-dose DHEA group.the expression level of Grem1 in cumulus cells in DHEA treatment group was lower than that in control group,and the reduction multiple of Grem1 in high dose group was greater than that in low dose group.The expression level of Ptgs2 was similar to that in Grem1,that is,the level of Ptgs2 in DHEA experimental group was lower than that in control group.In addition,the levels of Has2,Vcan and Ptgs1,the other markers related to embryo quality,were significantly up-regulated in the experimental group compared with the control group.Has2 was up-regulated most obviously.The differences were statistically significant.The expression level of Caspase3 in cumulus cells showed that DHEA treatment group significantly decreased compared with the experimental group.The down-regulation multiple in high-dose group was more obvious than that in low-dose group,which is 0.8 times in low-dose group and 0.3 times in high-dose group.The expression level of Bcl2 l10 mRNA in the experimental group was higher than that in the control group.The differences were statistically significant.Conclusion After DHEA treatment,the apoptosis of cumulus cells can be inhibited,the rate of heteromorphic eggs is decreased and the quality of oocytes can be improved.更多还原