The Effect of Melatonin on Mitochondrial Function and Autophagy in In Vitro Matured Oocytes of Aged Mice

Zahraa Nasheed Hamad Almohammed, Fatemeh Moghani-Ghoroghi, I. ragerdi-Kashani, R. Fathi, Leila Sadat Tahaei, Mohamad Naji, P. Pasbakhsh
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引用次数: 19

Abstract

Objective This study examined the in vitro effect of melatonin on the protein synthesis of mitochondria, as well as autophagy in matured oocytes of aged mice. Materials and Methods In this experimental study, germinal vesicles (GV) oocytes were collected from aged (with the age of six-months-old) and young mice (with age range of 6-8 weeks old) and then cultured in the in vitro culture medium (IVM) for 24 hours to each metaphase II (MII) oocytes and then supplemented with melatonin at a concentration of 10 μM. The culture medium of MII oocytes was devoid of melatonin. Afterward, the expression of the SIRT-1 and LC3 was assessed by immunocytochemistry. ATP-dependent luciferin-luciferase bioluminescence assay was employed for the measurement of the ATP contents. Intracellular reactive oxygen specious (ROS) was detected by DCFH-DA, and the total antioxidant capacity (TAC) level was determined by TAC assay. Results The expression of SIRT-1 and LC3, as well as the measurement of the ATP content, was significantly increased in oocytes treated with melatonin compared with the oocytes receiving no treatment. Moreover, TAC was considerably higher in melatonin-treated oocytes than oocytes receiving no treatment. On the other hand, the level of ROS was significantly decreased in oocytes treated with melatonin in comparison with the untreated oocytes. The results indicated that melatonin considerably improved the development of oocytes as well. Conclusion According to the data, melatonin increased mitochondrial function and autophagy via an increase in the expression of SIRT1 and LC3, as well as the ATP contents while it decreased the levels of ROS and increased TAC in oocytes derived from aged mice.
褪黑素对老年小鼠体外成熟卵母细胞线粒体功能和自噬的影响
目的研究褪黑素对衰老小鼠成熟卵母细胞线粒体蛋白合成及自噬的体外影响。材料与方法本实验取老龄小鼠(6月龄)和幼龄小鼠(6-8周龄)的生发囊泡(GV)卵母细胞,在体外培养基(IVM)中培养24小时至每个中期II (MII)卵母细胞,然后添加浓度为10 μM的褪黑素。MII卵母细胞培养基缺乏褪黑素。免疫细胞化学检测SIRT-1和LC3的表达。ATP依赖性荧光素-荧光素酶生物发光法测定ATP含量。DCFH-DA法检测细胞内活性氧(ROS)含量,TAC法测定细胞总抗氧化能力(TAC)水平。结果与未处理的卵母细胞相比,褪黑素处理的卵母细胞SIRT-1和LC3的表达以及ATP含量的测定明显增加。此外,褪黑激素处理的卵母细胞的TAC明显高于未处理的卵母细胞。另一方面,与未处理的卵母细胞相比,褪黑激素处理的卵母细胞的ROS水平显著降低。结果表明,褪黑素也显著改善卵母细胞的发育。结论褪黑素通过提高衰老小鼠卵母细胞中SIRT1和LC3的表达以及ATP含量来提高线粒体功能和自噬,同时降低ROS水平,增加TAC。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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