3种药物对老化卵母细胞4种母源mRNA水平的影响

The Effects of Three Drugs on Maternal mRNA Level in Aged Oocytes

利用Real-Time PCR技术定量分析了绵羊(Ovis aries)卵母细胞在生长-成熟-老化过程4种母源mRNA(Mater、Zar1、Dnmt1、GDF9)发生的动态变化,以及3种药物(咖啡因、DTT和矾酸盐)对老化卵母细胞母源mRNA水平的影响。结果显示,除Dnmt1在12 h开始下降外,卵母细胞中其他3个基因的表达量都是从生发泡期逐渐下降,且在体外培养至24 h降到最低,然后随着卵母细胞的老化又逐渐上升。其中GDF9和Mater 2个基因在30 h组的卵母细胞中的表达量显著高于24 h组卵母细胞(P<0.05)。30 h卵母细胞中Zar1和Dnmt1的转录水平与24 h卵母细胞差异不显著(P>0.05)。除Zar1外,咖啡因和DTT都显著降低了老化卵母细胞中其他3个基因的表达量(P<0.05),且所有基因表达量与24 h组卵母细胞差异不显著(P>0.05)。矾酸盐处理后的卵母细胞其4种母源mRNA水平都是最高,除Zar1外其他3个基因表达量都显著高于24 h组(P<0.05)。结论是,Mater、Zar1、Dnmt1、GDF9 4种母源mRNA水平与卵母细胞质量成反比,咖啡因和DTT能在一定程度上延缓卵母细胞的衰老,改善卵母细胞质量;而矾酸盐则加速了卵母细胞老化进程,降低了卵母细胞质量。

This study used Real-Time PCR technique to quantitatively analyze the expression of the maternal mRNA（Mater,Zar1,Dnmt1 and GDF9） in growth-maturation-aging process of ovine oocytes and investigated the effect of caffeine,DTT and vanadate on the level of maternal mRNA in aged oocytes,respectively.The results showed that the expression of GDF9,Mater and Zar1 decreased gradually from germinal vesicle phase to MⅡ phase,while the expression of Dnmt1 decreased gradually from 12 h of maturation culture.The level of all above maternal mRNAs was the lowest at MⅡ phase and increased with aging.The expression of Mater,Dnmt1 and GDF9 in caffeine treated-or DTT-treated aged oocytes was lower than that of aged oocytes（P 0.05）,but not different from that of matured oocytes（P 0.05）.The expression of Mater,Zar1,Dnmt1,and GDF9 in vanadate-treated oocytes was the highest and significantly higher than that of MII group except for Zar1（P 0.05）.Conclusion： the level of Mater,Zar1,Dnmt1,and GDF9 is in contrast with oocytes quality.Both caffeine and DTT delay aging progression to some extent and improve the oocyte quality.On the contrary,vanadate accelerates aging progression.