Hassani Roya, Asgari Hamid Reza, Koruji Morteza, Zandieh Zahra, Nazmara Zohreh
Department of Anatomy, School of Medicine, Iran University of Medical Sciences, Tehran, Iran.
Stem Cell and Regenerative Medicine Research Center, Iran University of Medical Sciences, Tehran, Iran.
J Assist Reprod Genet. 2025 Aug 18. doi: 10.1007/s10815-025-03597-0.
Cryopreservation can lead to sperm damage, which is why incorporating extracellular vesicles derived from normal semen can help protect against the damage caused by freezing.
Extracellular vesicles were obtained from normozoospermic semen samples (n = 15), and their effects on the cryopreservation process were studied in three groups: the control group (A, n = 6), the extracellular vesicles treated before freezing group (B, n = 6), and the extracellular vesicles treated after freezing group (C, n = 6). Characteristics of sperm after thawing, including morphology, viability, motility, total antioxidant capacity (TAC), the rate of apoptosis, mitochondrial membrane potential (MMP), and the DNA fragmentation index (DFI), as well as the expression levels of the BAX, BCL, CRISP, and KRAS genes, were examined.
Group B displayed a significant increase in total motility, progressive motility, normal morphology, and MMP compared to groups A, and DNA fragmentation (DFI) showed a significant decrease compared to groups A and C. Also, vitality and TAC were significantly increased in group B compared to groups A and C. Decreased level of apoptosis was evident in group B compared to group A. Group A exhibited a significant increase in the expression of BAX/BCL ratio compared to groups B and C. Significantly higher expression of KRAS and CRISP was evident in group B compared to groups A and C.
Extracellular vesicles obtained from normozoospermic seminal plasma could protect against the adverse cryopreservation effects through increased MMP and TAC, as well as decreased expression of BAX/BCL ratio and increased KRAS and CRISP.
冷冻保存会导致精子损伤，这就是为什么加入源自正常精液的细胞外囊泡有助于防止冷冻造成的损伤。
从正常精液样本（n = 15）中获取细胞外囊泡，并在三组中研究其对冷冻保存过程的影响：对照组（A组，n = 6）、冷冻前用细胞外囊泡处理组（B组，n = 6）和冷冻后用细胞外囊泡处理组（C组，n = 6）。检测解冻后精子的特征，包括形态、活力、运动能力、总抗氧化能力（TAC）、凋亡率、线粒体膜电位（MMP）和DNA碎片化指数（DFI），以及BAX、BCL、CRISP和KRAS基因的表达水平。
与A组相比，B组的总运动能力、前向运动能力、正常形态和MMP显著增加，与A组和C组相比，DNA碎片化（DFI）显著降低。此外，与A组和C组相比，B组的活力和TAC显著增加。与A组相比，B组的凋亡水平明显降低。与B组和C组相比，A组的BAX/BCL比值表达显著增加。与A组和C组相比，B组的KRAS和CRISP表达明显更高。
从正常精液血浆中获得的细胞外囊泡可以通过增加MMP和TAC，以及降低BAX/BCL比值的表达和增加KRAS和CRISP来防止冷冻保存的不良影响。
