Effects of cilostamide and forskolin on the meiotic resumption and embryonic development of immature human oocytes
Effects of cilostamide and forskolin on the meiotic resumption and embryonic development of immature human oocytes
复制标题
西洛酰胺和毛喉素对未成熟人卵母细胞减数分裂恢复和胚胎发育的影响。
DOI：
10.1093/humrep/dem344
复制
发表时间：
2008-03-01
影响因子：
6.1
通讯作者：
Wang, Ning-ning
中科院分区：
文献类型：
--
作者：
Shu, Yi-min;Zeng, Hai-tao;Wang, Ning-ning
背景：为了获得卵母细胞的胞质成熟，人们使用了PDE3特异性抑制剂西洛胺和腺苷环化酶激活剂Forsklin来延长未成熟卵母细胞的预成熟培养。方法：从未受刺激的卵巢获取卵丘-卵母细胞复合体，分别在20mU西洛胺或50mU福司可林单独或联合培养6、12、24、48h。在这些指定的时间点检测细胞间缝隙连接通讯(GJC)水平和成熟状态。两相培养54h后获得的中期II卵母细胞(0~24 h使用减数分裂抑制剂，24~54 h不使用减数分裂抑制剂)进行胞质内单精子注射，并继续培养5天。结果：与人未成熟卵母细胞连续培养后，西洛胺和Forsklin均可延缓自然减数分裂进程。在减数分裂抑制培养后6、12、24和48h，西洛胺和Forsklin联合处理组生发泡破裂(GVBD)率显著低于对照组(均P<0.05)。西洛胺和福司可林合用时，GJC的丢失也延迟了6h。西洛胺和福司可林联合处理的受精率显著高于对照。尽管卵母细胞成熟率和卵裂率相似，但西洛胺和Forsklin处理组的囊胚形成率略有增加，但差异不显著。结论：西洛胺和福司可林联合应用对防止GJC丢失和恢复减数分裂具有协同作用，对卵母细胞的发育能力有积极的影响。
BACKGROUND: In an attempt to allow for acquisition of oocyte cytoplasmic maturation, PDE3 specific inhibitor, cilostamide and adenylate cyclase activator, forskolin were used to extend pre-maturation culture of immature human oocytes. METHODS: Cumulus-oocyte complexes retrieved from unstimulated ovaries were continuously cultured under 20 mu M cilostamide or 50 mu M forskolin, alone or in combination for 6, 12, 24 or 48 h, respectively. Levels of intercellular gap junction communication (GJC) and maturational status were examined at these designated time points. Metaphase II oocytes obtained following 54 h biphasic culture (with meiotic inhibitors from 0 to 24 h, no meiotic inhibitors from 24 to 54 h) were subject to intracytoplasmic sperm injection and embryos were cultured for five more days. RESULTS: Both cilostamide and forskolin delayed spontaneous meiotic progression after continuous culture with immature human oocytes. Combined treatment of cilostamide and forskolin significantly lowered the rates of germinal vesicle breakdown (GVBD) at 6, 12, 24 or 48 h after meiotic inhibitory culture, when compared with the control (all P < 0.05). A delay of 6 h for the loss of GJC was also observed under the combined treatment of cilostamide and forskolin. The fertilization rate was significantly higher under the combined treatment of cilostamide and forskolin than that of the control. Although the rates of oocyte maturation and embryo cleavage were similar among groups, there was a slight but non-significant increase in blastocyst formation rate with the treatment of cilostamide and forskolin. CONCLUSIONS: Combined treatment of cilostamide and forskolin positively influences oocyte developmental competence by exhibiting a synergistic effect on the prevention of GJC loss and resumption of meiosis.