生殖医学杂志2017，Vol.26Issue(5)：470-476,7.DOI:10.3969/j.issn.1004-3845.2017.05.016
褪黑素对老化卵母细胞体外发育能力的改善效果评价
Effect of melatonin supplementation on improving developmental potential of aging oocytes in IVF
摘要
Abstract
Objective:To explore effect of supplementing melatonin into medium of in vitro culture on development potential of aging oocytes.Methods:The MII oocytes retrieved from ovaries were incubated with M2 culture medium without (as control) or with H2O2 at different concentrations (10,50,100,150 μmol/L) for 3 hours to induce oocyte aging.The 2-cell rate and blastocyst rate were counted after in vitro fertilization (IVF).The mitochondrial mass and activity (Mitotracker Red and JC-1),reactive oxygen species (ROS) levels,as well as mitochondrial DNA (mtDNA) copy number were also detected in control and aging oocytes.After treated with 100 μmol/L H2O2,the aging oocytes were in vitro fertilized and cultured in medium without or with melatonin at different concentrations (10-5,10-7,10-9 mol/L).Then,the 2-cell rate and blastocyst rate were counted,and the cell number of blastocysts and apoptotic rate were detected in the each groups.Results:Different concentrations of H2O2 induced aging of oocyte showed that the blastocyst development rate was decreased along with the increase of H2O2 concentration.The blastocyst rate of aging oocytes induced by 50 μmol/L and 100 μmol/L H2O2 was (26.27±0.06)% and (28.46±3.45)%,respectively,which were significantly lower than that in the control group [(34.90±1.77)%] (P<0.05).H2O2 at higher concentration (100 or 150 μmol/L) induced significantly higher intracellular reactive oxygen species (ROS) in aging oocytes (P<0.05),as well as a tendency of reduced mitochondrial mass and increased mitochondrial membrane potential (MMP),but the difference was not significant (P>0.05).The blastocyst rate [(29.42±2.39)% vs.(20.87±4.12)%,P<0.05) and the cell number of blastocysts from aging oocytes treated with 100 μmol/L H2O2 were significantly increased [(39.36±9.78) vs.(37.91±4.25),P<0.05) and the apoptotic rate was significantly decreased (2.57% vs.3.18%,P<0.05)by supplementing 10-9 mol/L melatonin into culture medium.And these indicators reached a similar level of development without aging treatment group.Conclusions:The lower development rate and development quality in aging oocytes in vitro fertilization can be reversed and improved by supplementing melatonin into culture medium.关键词
老化卵母细胞/褪黑素/线粒体/体外培养Key words
Aging oocyte/Melatonin/Mitochondria/In vitro culture引用本文复制引用
王伟周,任立坤,沈玉良,魏德莉,陈昭烈..褪黑素对老化卵母细胞体外发育能力的改善效果评价[J].生殖医学杂志,2017,26(5):470-476,7.基金项目
海军总医院创新培育基金(CXPY201408) （CXPY201408）
