Suppressionofmitochondrialoxidativephosphorylationoninvitrom

　　摘要： 通过建立氧化磷酸化（oxidative phosphorylation, OXPHOS）活性抑制模型，分析线粒体能量代谢在卵子成熟、受精和胚胎发育过程中的作用。 方法：卵子体外成熟培养液中引入10和100 nmol/L羰基氰4-(三氟甲氧基)苯腙［carbonyl cyanide p-(tri-fluromethoxy) phenyl-hydrazone，FCCP］，以特异性抑制线粒体OXPHOS活性，分析卵子线粒体膜电位强度、三磷酸腺苷含量以及线粒体DNA拷贝数，以验证FCCP效应；检测卵子的活性氧（reactive oxygen species，ROS）含量、纺缍体和染色体结构完整性，并统计分析卵子体外生发泡破裂（germinal vesicle breakdown，GVBD）率、核成熟率、受精率及囊胚形成率。 结果： FCCP(10和100 nmol/L)处理组与对照组相比，卵子的核成熟率（55.8 %和47.3%， vs. 62.9%）、纺锤体/染色体结构完整率（37.9% 和34.7%， vs. 61.9%）、ROS水平(0.67和 0.59， vs. 0.94)和囊胚形成率（57.9%和41.8%，vs. 68.3%）均有显著降低（P 0.05）。 结论：线粒体能量缺乏显著降低卵子的核成熟率、ROS生成、纺锤体/染色体结构完整率和囊胚形成率，但对卵子的GVBD发生率和受精率没有显著改变。

　　关键词:

　　线粒体,

　　氧化磷酸化,

　　卵母细胞,

　　受精, 体外

　　Abstract: To investigate the roles of mitochondrial oxidative phosphorylation (OXPHOS) capacity in oocyte maturation, fertilization and embryo development. Methods: Carbonyl cyanide p- (tri-fluromethoxy) phenyl-hydrazone (FCCP), a metabolic inhibitor of mitochondria, was introduced into culture medium. The integrity of spindle and chromosome alignment, reactive oxygen species (ROS) levels, and rates of maturation, germinal vesicle breakdown, fertilization and blastulation were assessed in vitro. Results: Significant decreases were detected in the percentages of oocytes with nuclear maturation, normal spindle formation and chromosome alignment, ROS levels and capable for blastocyst formation between oocytes treated with FCCP and non-treated (control group), 55.8%, 37.9%, 0.67 and 57.9% (FCCP 10 nmol/L group), 47.3%,34.7%, 0.59 and 41.8% (FCCP 100 nmol/L group) versus 62.9%, 61.9%,0.94 and 68.3% (control group) respectively, P < 0.05. However, No significant differences were found in the rates of GVBD and fertilization in oocytes from the FCCP treated and the control. Conclusion: Inhibition of mitochondrial metabolic capacity resulted in decreased the percentages of oocytes with nuclear maturation, normal spindle formation and chromosome alignment, ROS levels and capable for blastocyst formation. But the treatment of FCCP did not affect the rate of fertilization.

　　Key words:

　　Mitochondria,

　　Oxidative phosphorylation,

　　Oocytes,

　　Fertilization in vitro

　　引用本文

　　葛红山，李晓和，张帆，陈华，习海涛，黄检英，朱春芳，吕杰强. 抑制线粒体氧化磷酸化活性对卵子体外成熟、受精及发育的影响[J]. 北京大学学报（医学版）, 2013, 45(6): 864-868.

　　GE Hong-shan, LI Xiao-he, ZHANG Fan, CHEN Hua, XI Hai-tao, HUANG Jian-ying, ZHU Chun-fang, LV Jie-qiang. Suppression of mitochondrial oxidative phosphorylation on in vitro maturation, fertilization and developmental competence of oocytes[J]. Journal of Peking University(Health Sciences), 2013, 45(6): 864-868.

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