Department of Obstetrics and Gynecology, Graduate School of Medicine, Mie University, 2-174 Edo-bashi, Tsu 514-8507, Japan.
Center of Advanced Reproductive Medicine, Mie University Hospital, 2-174 Edobashi, Tsu 514-8507, Japan.
Cells. 2023 Nov 7;12(22):2585. doi: 10.3390/cells12222585.
Sperm DNA fragmentation (SDF) that occurs during the freezing-thawing of sperm may negatively impact the treatment outcomes of assisted reproductive technologies (ART). In a previous study, we developed a human sperm cryopreservation reagent containing carboxylated poly-L-lysine (CPLL) that reduced SDF after freeze-thawing compared with clinically popular cryopreservation reagents containing human serum albumin. However, it is unclear whether CPLL reduces SDF, as it differed from the constituents of the commercial cryopreservation reagents used for comparison. Therefore, here, we examined whether CPLL reduces the SDF of human sperm and evaluated reactive oxygen species (ROS) levels and lipid peroxidation (LPO), which are the causes of SDF; mitochondrial injury, ROS production; and impaired sperm motility. Furthermore, optimal antioxidants and their concentrations that could further enhance the reduction in SDF were determined for future clinical application in ART and underwent the same functional evaluations. CPLL can reduce SDF via inhibition of intracytoplasmic ROS and LPO. Furthermore, the addition of 0.1 mM resveratrol avoided the enhancement of SDF, which potentially affects mitochondrial and cytoplasmic ROS and LPO. This novel human sperm cryopreservation reagent containing CPLL and resveratrol has the potential to improve treatment outcomes in ART using frozen sperm.
精子 DNA 碎片化(SDF)在精子冷冻-解冻过程中发生,可能会对辅助生殖技术(ART)的治疗结果产生负面影响。在之前的研究中,我们开发了一种含有聚赖氨酸(CPLL)的人类精子冷冻保存试剂,与含有人血清白蛋白的临床流行的冷冻保存试剂相比,该试剂可减少冷冻-解冻后的 SDF。然而,目前尚不清楚 CPLL 是否会降低 SDF,因为它与用于比较的商业冷冻保存试剂的成分不同。因此,在这里,我们检查了 CPLL 是否会降低人精子的 SDF,并评估了活性氧(ROS)水平和脂质过氧化(LPO),这是 SDF 的原因;线粒体损伤、ROS 产生;以及受损的精子运动。此外,确定了最佳抗氧化剂及其浓度,以便在未来的 ART 临床应用中进一步增强 SDF 的降低,并进行了相同的功能评估。CPLL 可以通过抑制细胞内 ROS 和 LPO 来降低 SDF。此外,添加 0.1mM 白藜芦醇可避免增强 SDF,这可能会影响线粒体和细胞质 ROS 和 LPO。这种含有 CPLL 和白藜芦醇的新型人类精子冷冻保存试剂有可能改善使用冷冻精子的 ART 的治疗结果。