| 一氧化氮对小鼠卵母细胞自发成熟的影响及作用机制 |
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| 引用本文： | 王松波,周波,张美佳,夏国良.一氧化氮对小鼠卵母细胞自发成熟的影响及作用机制[J].农业生物技术学报,2009,17(5):797-801. |
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| 作者姓名： | 王松波 周波 张美佳 夏国良 |
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| 作者单位： | 1. 中国农业大学生物学院,北京,100193;华南农业大学动物科学学院,广州,510642
2. 中国农业大学生物学院,北京,100193 |
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| 基金项目： | 国家自然科学基金项目，国家重大基础研究发展规划(973)项目 |
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| 摘 要： | 为研究一氧化氮（NO）供体硝普钠（nitroprusside sodium，SNP）对小鼠卵母细胞体外自发成熟的作用，并进一步探讨NO影响卵母细胞体外自发成熟的可能机制，我们采用了体外细胞培养方法和放射免疫分析法（RIA）。细胞培养研究结果显示，NO供体SNP（1mM）能够延迟卵丘卵母细胞复合体(cumulus-oocyte complexes, COCs) 自发成熟过程中GVBD的发生，抑制PB1的释放。放射免疫分析（RIA）实验结果表明，1mM SNP能够显著的升高COCs内cGMP的水平，而可溶性鸟苷酸环化酶（sGC）抑制剂ODQ（10μM）能够消除SNP对cGMP水平升高的促进作用。同时10μM ODQ还能够逆转SNP对卵母细胞自发成熟的抑制作用，而PKG抑制剂KT5823（1μM）却不能够逆转SNP对COCs自发成熟的抑制作用。以上结果说明NO是通过激活sGC，提高细胞内cGMP水平来发挥其作用的，但cGMP下游的PKG信号通路并不参与NO调控的COCs自发成熟过程。
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| 关 键 词： | 一氧化氮 硝普钠 小鼠卵母细胞 自发成熟 |
| 收稿时间： | 2008-10-27 |
| 修稿时间： | 2008-12-3 |
Effects of Nitric Oxide on Mouse Oocyte Spontaneous Maturation and Its Signal Pathway |
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| Abstract: | The objective of the present study was to investigate the effects of NO donor nitroprusside sodium (SNP) (1mM) on mouse oocytes spontaneous maturation and to explore the mechanisms by which NO influenced oocyte spontaneous maturation. Oocytes maturation was evaluated through in vitro cell culture and intracellular cGMP levels were determined by Radioimmunoassay (RIA). The results showed that NO donor SNP (1mM) could delay the occurrence of GVBD and inhibit the first polar body (PB1) extrusion of cumulus-oocyte complexes (COCs). The data of RIA demonstrated that NO donor SNP significantly increased the cGMP levels in COCs and that the stimulative effect of SNP on cGMP level could be eliminated by soluble guanylate cyclase (sGC) inhibitor ODQ (10μM). ODQ could also reverse the inhibitory effects of SNP on COCs spontaneous maturation. However, the suppressive effects of SNP on COCs spontaneous maturation were not abolished by PKG inhibitor KT5823 (1μM). These results indicated that NO elicited its effects on spontaneous maturation through activating sGC and elevating the intracellular cGMP levels and suggested that PKG signal pathway was not involved in NO-mediated mouse COCs spontaneous maturation. |
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