外源褪黑素对牛卵母细胞体外成熟的影响  

Effect of Exogenous Melatonin(MT) in Bovine Oocyte in vitro Maturation

作　　者：高超[1] 田秀芝[1] 张璐[1] 徐静[1] 汪锋[1] 卓志勇[1] 戴蕴平[1] 刘国世[1] 

机构地区：[1]中国农业大学动物科技学院,北京100391

出　　处：《中国农业科学》2011年第17期3634-3640,共7页Scientia Agricultura Sinica

基　　金：国家"863"计划项目(2008AA1011007);农业部转基因重大专项(2009ZX0800-020B;2008ZX08008-005);国家科技支撑计划项目(2011BAD19B02-4)

摘　　要：【目的】探索外源褪黑素(melatonin,MT)处理对牛卵母细胞体外成熟的影响从而优化牛体外胚胎生产体系。【方法】在牛卵母细胞成熟过程中添加不同浓度的MT,观察并统计其体外受精后胚胎的卵裂率、囊胚率和囊胚细胞数。应用放射免疫分析法检测血清、卵泡液和成熟培养液内MT与孕酮(P)、雌激素(E2)、促卵泡素(FSH)、促黄体生成素(LH)水平的变化并检测卵母细胞的活力和活性氧(ROS)的浓度。【结果】存在于牛卵泡液中的MT浓度与血清中的相近,添加10-11 mol.L-1 MT组的P浓度(2.01±0.33)ng.mL-1显著高于对照组((0.87±0.10)ng.mL-1,P<0.05)。与对照组((67.32±7.30)%)相比,10-9 mol.L-1 MT组卵裂率显著提高((77.81±7.06)%,P<0.05),并能提高囊胚率且囊胚细胞数((29.61±9.55)%,(90.30±28.74)个),显著高于对照组((19.64±9.22)%,(75.07±25.98)个,P<0.05);同时,10-9 mol.L-1 MT组卵母细胞的活力高于对照组,并能降低卵母细胞内的ROS含量但没有显著差异(P>0.05)。【结论】牛卵泡液中存在MT,适当浓度的外源MT能促进牛卵母细胞体外成熟。[Objective] To optimize the bovine embryo in vitro production system, the effect of exogenous melatonin （MT） on improvement of bovine oocyte in vitro maturation was studied. [ Method] Immature oocytes were cultured in vitro inmaturation medium supplemented with different concentrations of melatonin, then IVF statistical analysis of embryo cleavage rate, blastocyst rate and blastocyst cell number was conducted. Hormone levels were detected by radioimmunoassay in follicular fluid and the maturation medium and the the oocyte viability and reactive oxygen （ROS） levels were determined. [Result] MT existed in bovine follicular fluid and similar with serum. In 10-11mol·L^-1 treatment, the progesterone （P） levels （2.01±0.33） ng·mL^-1 （P〈0.05） of maturation medium was significantly higher than that of controls （0.87i0.10） ng·mL^-1 （P〈 0.05）. Compared with the control group （67.32±7.30）%, cleavage rate in 10^-9mol·L^-1 MT treatment （77.81±7.06）% was significantly increased and the blastocyst rate and cell number of blastocyst （29.61±9.55）%, （90.30~28.74） were significantly higher than that of controls （19.64±9.22）%, （75.074-25.98） （P〈0.05）. 10^-9mol·L^-1 MT treatment improved the oocyte viability and decreased the ROS level （P〉0.05）. [Conclusion] Melatonin exists in bovine follicular fluid and exogenous melatonin at a proper concentration can improve the in vitro maturation of bovine oocytes.

关 键 词：褪黑素 卵母细胞 体外成熟 活性氧 牛 

分 类 号：S823[农业科学—畜牧学]