体外成熟技术及补充α-硫辛酸对卵母细胞成熟率的影响：聚焦于卵母细胞的氧化状态
Effect of In Vitro Maturation Technique and Alpha Lipoic Acid Supplementation on Oocyte Maturation Rate: Focus on Oxidative Status of Oocytes.
作者信息
Zavareh Saeed, Karimi Isaac, Salehnia Mojdeh, Rahnama Ali
机构信息
School of Biology, Damghan University, Damghan, Iran; Institute of Biological Sciences, Damghan University, Damghan, Iran.
Laboratory of Molecular and Cellular Biology, Department of Basic Veterinary Sciences, School of Veterinary Medicine, Razi University, Kermanshah, Iran.
出版信息
Int J Fertil Steril. 2016 Jan-Mar;9(4):442-51. doi: 10.22074/ijfs.2015.4601. Epub 2015 Dec 23.
BACKGROUND
Therapeutic potential of in vitro maturation (IVM) in infertility is growing with great promise. Although significant progress is obtained in recent years, existing IVM protocols are far from favorable results. The first aim of this study was to investigate whether two step IVM manner change reactive oxygen species (ROS) and total anti- oxidant capacity (TAC) levels. The second aim was to find the effect of alpha lipoic acid (ALA) supplementation on oocyte maturation rate and on ROS/TAC levels during IVM.
MATERIALS AND METHODS
In this experimental study, mouse germinal vesicle (GV) oocytes divided into cumulus denuded oocytes (DOs) and cumulus oocyte complexes (COCs) groups. GVs were matured in vitro in the presence or absence of ALA only for 18 hours (control) or with pre-culture of forskolin plus cilostamide for an additional 18 hours. Matured oocytes obtained following 18 and 36 hours based on experimental design. In parallel, the ROS and TAC levels were measured at different time (0, 18 and 36 hours) by 2',7'-dichlorodihydrofluorescein (DCFH) probe and ferric reducing/antioxidant power (FRAP) assay, respectively.
RESULTS
Maturation rate of COCs was significantly higher than DOs in control group (P<0.05), while there was no significant difference between COCs and DOs when were pre-cultured with forskolin plus cilostamide. ROS and TAC levels was increased and decreased respectively in DOs after 18 hours while in COCs did not change at 18 hours and showed a significant increase and decrease respectively at 36 hours (P<0.05). ROS and TAC levels in the presence of ALA were significantly decreased and increased respectively after 36 hours (P<0.05) whereas, maturation rates of COCs and DOs were similar to their corresponding control groups.
CONCLUSION
ALA decreased ROS and increased TAC but could not affect maturation rate of both COCs and DOs in one or two step IVM manner.
背景
体外成熟（IVM）在不孕症治疗中的潜力不断增长，前景广阔。尽管近年来取得了显著进展，但现有的IVM方案远未取得理想效果。本研究的首要目的是探讨两步IVM方式是否会改变活性氧（ROS）和总抗氧化能力（TAC）水平。第二个目的是找出补充α-硫辛酸（ALA）对IVM期间卵母细胞成熟率以及ROS/TAC水平的影响。
材料与方法
在本实验研究中，将小鼠生发泡（GV）卵母细胞分为裸卵（DOs）和卵丘卵母细胞复合体（COCs）组。GVs在有或无ALA的情况下仅体外成熟18小时（对照组），或先用福斯高林加西洛他唑预培养18小时，再继续培养18小时。根据实验设计，在18小时和36小时后获得成熟卵母细胞。同时，分别在不同时间点（0、18和36小时）通过2',7'-二氯二氢荧光素（DCFH）探针和铁还原/抗氧化能力（FRAP）测定法测量ROS和TAC水平。
结果
对照组中COCs的成熟率显著高于DOs（P<0.05），而在用福斯高林加西洛他唑预培养时，COCs和DOs之间没有显著差异。DOs在18小时后ROS水平升高，TAC水平降低，而COCs在18小时时未发生变化，在36小时时分别显著升高和降低（P<0.05）。在添加ALA的情况下，36小时后ROS水平显著降低，TAC水平显著升高（P<0.05），而COCs和DOs的成熟率与其相应对照组相似。
结论
ALA降低了ROS水平，提高了TAC水平，但在一步或两步IVM方式下均不影响COCs和DOs的成熟率。
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