Moghimi Khorasgani Ali, Moradi Reza, Jafarpour Farnoosh, Ghazvinizadehgan Faezeh, Ostadhosseini Somayyeh, Heydarnezhad Alireza, Fouladi-Nashta Ali Akbar, Nasr-Esfahani Mohammad Hossein
Department of Reproductive Biotechnology, Reproductive Biomedicine Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran.
Department of Agricultural Biotechnology, College of Agriculture, Isfahan University of Technology, Isfahan, Iran.
Cell J. 2021 Jul;23(2):164-173. doi: 10.22074/cellj.2021.7071. Epub 2021 May 26.
Alpha-lipoic acid (ALA) as a strong antioxidant has a protective effect. This study was designed to assess whether supplementation of maturation medium with ALA during maturation (IVM) can attenuate the toxic effect of ethanol.
In this experimental study, to assess the antioxidant capacity of ALA challenged by 1% ethanol during maturation, immature ovine oocytes were exposed to 1% alcohol in the presence or absence of 25 μM ALA during oocyte maturation. The cumulus expansion index, intracellular reactive oxygen species (ROS), and thiol content levels were assessed in matured oocytes of various treatment groups. Consequently, the blastocyst formation rate of matured oocytes in various treatment groups were assessed. In addition, total cell number (TCN), cell allocation, DNA fragmentation, and relative gene expression of interested genes were assessed in resultant blastocysts.
The results revealed that alcohol significantly reduced cumulus cells (CCs) expansion index and blastocyst yield and rate of apoptosis in resultant embryos. Addition of 25 μM ALA to 1% ethanol during oocyte maturation decreased ROS level and elevated Thiolcontent. Furthermore, supplementation of maturation medium with ALA attenuated the effect of 1% ethanol and significantly increased the blastocyst formation and hatching rate as compared to control and ethanol groups. In addition, the quality of blastocysts produced in ALA+ethanol was improved based on the low number of TUNEL positive cells, the increased expression level of mRNA for pluripotency, and anti-oxidant markers, and decreased expression of apoptotic genes.
The current findings demonstrate that ALA can diminish the effect of ethanol, possibly by decreasing the ROS level and increasing Thiolcontent during oocyte maturation. Using the ALA supplement may have implications in protecting oocytes from alcohol toxicity in affected patients.
α-硫辛酸（ALA）作为一种强效抗氧化剂具有保护作用。本研究旨在评估在体外成熟（IVM）过程中向成熟培养基中添加ALA是否能减轻乙醇的毒性作用。
在本实验研究中，为评估在成熟过程中受到1%乙醇挑战的ALA的抗氧化能力，将未成熟的绵羊卵母细胞在卵母细胞成熟期间暴露于含或不含25μM ALA的1%酒精中。评估各处理组成熟卵母细胞的卵丘扩展指数、细胞内活性氧（ROS）和硫醇含量水平。随后，评估各处理组成熟卵母细胞的囊胚形成率。此外，评估所得囊胚的总细胞数（TCN）、细胞分配、DNA片段化以及感兴趣基因的相对基因表达。
结果显示，酒精显著降低了卵丘细胞（CCs）扩展指数、囊胚产量以及所得胚胎的凋亡率。在卵母细胞成熟期间向1%乙醇中添加25μM ALA可降低ROS水平并提高硫醇含量。此外，与对照组和乙醇组相比，在成熟培养基中添加ALA可减轻1%乙醇的影响，并显著提高囊胚形成率和孵化率。此外，基于TUNEL阳性细胞数量减少、多能性和抗氧化标志物mRNA表达水平增加以及凋亡基因表达降低，ALA + 乙醇组产生的囊胚质量得到改善。
当前研究结果表明，ALA可能通过在卵母细胞成熟期间降低ROS水平和增加硫醇含量来减轻乙醇的影响。使用ALA补充剂可能对保护受影响患者的卵母细胞免受酒精毒性具有重要意义。
