Faggi Melina, Paparella Cecilia, Perfumo Patricia, Teijeiro Juan Manuel
Laboratorio de Medicina Reproductiva. Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Rosario, Santa Fe, Argentina.
Unidad de Reproducción Humana Médicamente Asistida, Hospital del Centenario, Rosario, Santa Fe, Argentina.
J Assist Reprod Genet. 2025 Jan;42(1):335-342. doi: 10.1007/s10815-024-03328-x. Epub 2024 Nov 27.
Zinc is known to influence chromatin stability, motility and protection against oxidative stress. While swim-up remains the preferred method for selecting sperm in Assisted Reproductive Technologies (ART), concerns arise regarding sperm DNA fragmentation associated with this procedure. Given zinc's significant role in protecting sperm DNA integrity and motility, we aimed to investigate the impact of zinc supplementation during the swim-up process on sperm quality.
Semen samples from 203 normozoospermic men were used. Samples were divided into fractions and swim-up procedure was applied using human tubal fluid (mHTF) supplemented with three different concentrations of zinc or medium without supplementation as control. DNA fragmentation, chromatin maturity, reactive oxygen species (ROS) levels, motility and protein phosphorylation levels analyses were addressed to each fraction.
The sperm DNA fragmentation was reduced in sperm recovered by swim-up in media with all concentrations of zinc assayed with respect to the control (p < 0.0001). Aniline blue staining showed better chromatin maturity in sperm recovered with 2.5- and 3.5-mM zinc (p = 0.045; p = 0.021). Kinematic parameters such as curvilinear velocity and beat-cross frequency showed improvement with 2.5 mM zinc (p = 0.0080 and p = 0.0400), whereas straightness, linearity, and hypermotility showed improvement with 5 mM zinc (p = 0.0075, p = 0.0069, and p = 0.0244). Protein phosphorylation patterns showed changes associated with treatment with zinc, and only 5 mM zinc treatment showed a decrease in ROS levels.
The addition of zinc to mHTF provided optimal physiological conditions for sperm recovered through swim-up. This supplementation should be considered for selecting sperm for use in ART.
已知锌会影响染色质稳定性、活力以及对氧化应激的保护作用。在辅助生殖技术（ART）中，上游法仍是优选的精子筛选方法，但人们对该过程中与精子DNA片段化相关的问题表示担忧。鉴于锌在保护精子DNA完整性和活力方面的重要作用，我们旨在研究上游过程中补充锌对精子质量的影响。
使用来自203名正常精子男性的精液样本。将样本分成几份，采用添加三种不同浓度锌的人输卵管液（mHTF）或不添加锌的培养基作为对照进行上游处理。对每份样本进行DNA片段化、染色质成熟度、活性氧（ROS）水平、活力和蛋白质磷酸化水平分析。
与对照组相比，在所有测定浓度锌的培养基中通过上游法回收的精子中，精子DNA片段化减少（p < 0.0001）。苯胺蓝染色显示，在添加2.5 mM和3.5 mM锌回收的精子中染色质成熟度更好（p = 0.045；p = 0.021）。曲线速度和拍击交叉频率等运动学参数在添加2.5 mM锌时有所改善（p = 0.0080和p = 0.0400），而直线性、线性度和超活力在添加5 mM锌时有所改善（p = 0.0075，p = 0.0069和p = 0.0244）。蛋白质磷酸化模式显示与锌处理有关的变化，只有5 mM锌处理显示ROS水平降低。
在mHTF中添加锌为通过上游法回收的精子提供了最佳生理条件。在ART中选择精子时应考虑这种补充方法。
