目的:本文采用体外培养技术观察原癌基因c-myb对孕酮诱导的生发泡(GV)期小鼠裸卵体外成熟的影响。方法:建立小鼠裸卵体外培养模型,用不同浓度反义C-myb寡脱氧核苷酸(c-myb ASODNs)与GV期小鼠裸卵共孵育观察其对孕酮诱导的小鼠裸卵体外成熟的影响并探讨其机制。结果:在M199培养液中体外培养GV期小鼠裸卵24h,10μmol/L孕酮组与5μmol/L孕酮组比较有显著性差别(2 h GVBD% P〈0.05,8 h PBI% P〈0.05).与20μmol/L孕酮组比较无显著性差别。10μmol/L c-myb AFODNs能抑制孕酮(10μmol/L)诱导的小鼠裸卵体外成熟(2 h GVBD% P〈0.05,8 h PBI% P〈0.01)。1×10^-4μmol/L dbcAMP、10μg/ml肝素钠可分别单独抑制孕酮诱导的GV期小鼠卵母细胞体外成熟(2 h GVBD%均P〈0.01,8 h PBI%均P〈0.01).也可和反义c-myb ODN协同抑制孕酮诱导的卵母细胞体外成熟(2 h GVBD%均P〈0.01,8 h PBI%均P〈0.01)。结论:孕酮、原癌基因c-myb和cAMP、Ca^2+参与了GV期小鼠卵母细胞的体外成熟,孕酮、cAMP和Ca^2+调控卵母细胞成熟的机理可能与原癌基因c-myb表达有关。
To investigate the effects of c-myb on progesterone-induced mouse germinal vesicle(GV) stage denuded oocyte(DO) maturation in uitro. Methodsds: We used mouse GV stage oocyte cultured with special concentration progesterone, or/and antisense c-myb ODN, or/and db-cAMP, or/and heparin for 24 h, and observed oocyte maturation and analysed the relationship among them. Results: We cultured DO in the medium 199 for 24 h,and found 10μmol/L progesterone had morn significant effect than 5 μmol/L progesterone(2 h GVBD% P〈0.05, 8 h PBI% P〈0.05), but had not more significant effect than 20μmol/L prngesterone. We found that 16μmol/L antisense c-myb ODN significantly inhibited progesterone(10μmol/L)-induced mouse germinal vesicle stage oocyte maturation in uitro(2 h GVBD% P〈0.05,8 h PBI% P〈0.01 ). 1×10^-4 μmol/L dbcAMP, 100μg/ml heparin could single significantly inhibited progesterone-induced mouse GV stage oocyte maturation in uitro(2 h PBI% all P〈0.01,8 h PBI% all P〈0.01), and could enhanced the inhibition of 16 μmol/L antisense c-myb ODN(2 h GVBD% all P〈0.01,8h PBI% all P〈0.01). Conclusion: Progesterone, progesterone c-myb, cAMP and caldum all pay important role in regulating oocyte maturation and the mechanism of progesterone, cAMP and calcium in regulating oocyte maturation may be through the expression of pro-toonoogene c-myb.