评价米托醌对人精子冻融后功能参数、DNA 结构和与凋亡及抗氧化相关基因表达的影响。
Evaluation of the effect of mitoquinone on functional parameters, DNA structure, and genes expression related to the apoptotic and antioxidants of human sperm after freezing-thawing.
机构信息
Department of Veterinary Sciences, Garmsar Branch, Islamic Azad University, Garmsar, Iran.
Department of Biology, Islamic Azad University of Hamedan, Hamadan, Iran.
出版信息
Mol Biol Rep. 2024 Jan 23;51(1):183. doi: 10.1007/s11033-023-09020-0.
OBJECTIVE
Sperm freezing is considered as an effective way in assisted reproductive technology (ART) programs, it has detrimental effects on sperm function, due to the production of reactive oxygen species (ROS). This study aimed to investigate the potential of Mitoquinone (MitoQ) in inhibiting the production of mitochondrial ROS during sperm freezing.
METHODS
A total of 20 human normozoosperm samples were collected for this study. The samples were divided into four groups, each containing different concentrations of MitoQ (0, 0.2, 2, and 20 nM), and then subjected to the freezing process. After thawing, the sperm suspensions were evaluated for parameters including motility, morphology, acrosome integrity, adenosine triphosphate (ATP) level, intracellular ROS, viability, chromatin packaging, DNA denaturation, DNA fragmentation, as well as the expression of antioxidants (GPX, SOD) and apoptotic (Bax, Bcl) genes.
RESULTS
The results showed that total and progressive mobility of sperms significantly increased in the 2 nM group, while significantly decreased in the 20 nM group (p ≤ 0.05). Sperm morphology did not significantly improve across all the tested concentrations (p ≥ 0.05). Intracellular ROS levels showed a significant decrease and increase in the concentrations of 2 and 20 nM, respectively (p ≤ 0.05). Furthermore, a significant increase was observed in viability, ATP, acrosome integrity, chromatin packaging, and non-denatured and non-fragmented DNA after treatment with 2 nM of MitoQ, compared with the control group (p ≤ 0.05). Regarding gene expressions, the relative expressions of oxidative stress genes were increased in the 2 nM group and decreased in the 20 nM group (p ≤ 0.05), while no significant difference was observed in the expressions of apoptotic genes compared with the control group (p ≥ 0.05). All the comparisons were made with respect to the control group.
CONCLUSION
Adding the optimal concentration of MitoQ (2 nM) to the sperm freezing medium not only improves sperm functional parameters and reduces DNA damages, but also stimulates the expression of antioxidant genes, leading to even greater benefits for sperm cryopreservation.
目的
精子冷冻被认为是辅助生殖技术（ART）项目中的一种有效方法，但由于活性氧（ROS）的产生，它会对精子功能产生不利影响。本研究旨在探讨 mitoquinone（MitoQ）在抑制精子冷冻过程中线粒体 ROS 产生方面的潜力。
方法
本研究共收集了 20 个人正常精子样本。将这些样本分为四组，每组含有不同浓度的 MitoQ（0、0.2、2 和 20 nM），然后进行冷冻过程。解冻后，评估精子悬液的运动、形态、顶体完整性、三磷酸腺苷（ATP）水平、细胞内 ROS、活力、染色质包装、DNA 变性、DNA 碎片化，以及抗氧化剂（GPX、SOD）和凋亡（Bax、Bcl）基因的表达。
结果
结果显示，2 nM 组的总运动和渐进运动显著增加，而 20 nM 组显著降低（p≤0.05）。所有测试浓度的精子形态均无显著改善（p≥0.05）。细胞内 ROS 水平在 2 和 20 nM 浓度下分别显著降低和升高（p≤0.05）。此外，与对照组相比，用 2 nM MitoQ 处理后，活力、ATP、顶体完整性、染色质包装、非变性和非碎片化 DNA 显著增加（p≤0.05）。关于基因表达，氧化应激基因的相对表达在 2 nM 组中增加，在 20 nM 组中减少（p≤0.05），而与对照组相比，凋亡基因的表达没有显著差异（p≥0.05）。所有比较均与对照组进行。
结论
向精子冷冻培养基中添加最佳浓度的 MitoQ（2 nM）不仅可以改善精子功能参数，减少 DNA 损伤，还可以刺激抗氧化基因的表达，从而使精子冷冻保存更具优势。
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