一氧化氮对小鼠卵母细胞自发成熟的影响及作用通路  

Effects of Nitric Oxide on Mouse Oocyte Spontaneous Maturation and Its Signal Pathway

作　　者：王松波[1,2] 周波[1] 张美佳[1] 夏国良[1] 

机构地区：[1]中国农业大学生物学院,北京100193 [2]华南农业大学动物科学学院,广州510642

出　　处：《农业生物技术学报》2009年第5期797-801,共5页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金项目(No.30671508);国家重大基础研究发展规划(973)项目(No.2006CB504003和No.2007CB947401)资助

摘　　要：采用体外细胞培养和放射免疫分析法(radioimmunoassay,RIA)研究了一氧化氮(nitric oxide,NO)供体硝普钠(nitro-prusside sodium,SNP)对昆明小鼠(Mus musculus)卵母细胞体外自发成熟的影响及作用通路。结果表明,NO供体SNP(1mmol/L)能够延迟卵丘卵母细胞复合体(cumulus-oocyte complexes,COCs)自发成熟过程中生发泡破裂(germinal vesicle break-down,GVBD)的发生,抑制第1极体(the first polar body,PB1)的释放。1mmol/L SNP还能够显著提升COCs内环鸟苷一磷酸(cyclic GMP,cGMP)的水平,而可溶性鸟苷酸环化酶(soluble guanylate cyclase,sGC)抑制剂1H-[1,2,4]oxadiazolo[4,3-a]quinox-alin-1-one(ODQ()1μmol/L)能够消除SNP对cGMP水平的提升作用。同时ODQ还能够逆转SNP对卵母细胞自发成熟的抑制作用,而蛋白激酶G(protein kinase G,PKG)抑制剂KT5823(1μmol/L)却不能够逆转SNP对COCs自发成熟的抑制作用。实验结果说明NO是通过激活sGC,提高细胞内cGMP水平而发挥其对小鼠卵母细胞自发成熟作用的,但cGMP下游的PKG信号通路并不参与这一过程。The effects of nitric oxide（NO） donor nitroprusside sodium（SNP） on Kunming mouse（Mus musculus） oocyte sponta-neous maturation and its signal pathway were investigated by in vitro cell culture and radioimmunoassay（RIA）.The results showed that NO donor SNP（1 mmol/L） could delay the occurrence of germinal vesicle breakdown（GVBD） and inhibit the first polar body（PB1） extrusion of cumulus-oocyte complexes（COCs）.Meanwhile, NO donor SNP significantly increased the cyclic GMP（cGMP） levels in COCs and the stimulative effect of SNP on cGMP level could be eliminated by soluble guanylate cyclase（sGC） inhibitor 1H-[1,2,4]oxadiazolo [4,3-a]quinoxalin-1-one（ODQ）.Furthermore, ODQ could reverse the inhibitory effects of SNP on COCs sponta-neous maturation.However, the suppressive effects of SNP on COCs spontaneous maturation were not abolished by protein kinase G（PKG） inhibitor KT5823（1μmol/L）.Experiment findings indicate that NO elicits its effects on mouse oocyte spontaneous maturation through activating sGC and elevating the intracellular cGMP levels and suggest that PKG signal pathway is not involved in this process.

关 键 词：一氧化氮 硝普钠 小鼠卵母细胞 自发成熟 

分 类 号：S188[农业科学—农业基础科学]