Galarza Diego A, Duma Mauricio, Carreño-Aguilar Karla, Sañay-Sañay José, Bernardi Cristina, Soria Manuel E, Méndez María S
Laboratorio de Biotecnología de la Reproducción Animal, Facultad de Ciencias Agropecuarias, Universidad de Cuenca, Cuenca EC010205, Ecuador; Centro Latinoamericano de Formación de Especies Mayores y Menores, CLAFEM, Cuenca, Ecuador.
Laboratorio de Biotecnología de la Reproducción Animal, Facultad de Ciencias Agropecuarias, Universidad de Cuenca, Cuenca EC010205, Ecuador.
Anim Reprod Sci. 2025 Aug;279:107886. doi: 10.1016/j.anireprosci.2025.107886. Epub 2025 Jun 9.
Resveratrol (RES) is a powerful antioxidant that has recently been shown to improve sperm survival subjected to different cryopreservation methods. This study evaluated the effect of RES supplementation in conventional slow (CS) freezing and kinetic vitrification (VIT) media on dog sperm quality variables. Twenty pooled sperm samples from 30 orchiectomized adult dogs were used (three epididymal samples / pool). In each pooled sample, four treatments were established based on RES supplementation (control [no added] or 100 µM) and the cryopreservation method (CS or VIT): CS-Control, CS+RES, VIT-Control and VIT+RES. The CS freezing was performed by exposing sperm samples in 0.25 ml straws to liquid nitrogen (LN₂) vapors. Vitrification was made by directly submerging 30 µl sperm droplets into LN₂. The results initially demonstrated that VIT treatments produced outcomes similar to those of CS freezing in terms of sperm kinematics and membrane integrity. Sperm samples from the CS+RES treatment showed higher percentages (P < 0.05) of motility, viability, acrosomal integrity, and DNA integrity compared to those from the CS-Control treatment. Furthermore, both CS+RES and VIT+RES treatments obtained a lower percentage (P < 0.05) of sperm DNA fragmentation compared to both CS-Control and VIT-Control treatments. Despite this, RES negatively affected certain kinematic parameters (e.g., velocities, straightness, and beat cross frequency) and sperm head dimensions after vitrification. In conclusion, resveratrol improved sperm motility, viability, and acrosomal integrity and reduced DNA fragmentation following conventional slow freezing. While it also reduced DNA fragmentation after vitrification, it negatively affected sperm kinematics in dog epididymal sperm.
白藜芦醇（RES）是一种强大的抗氧化剂，最近的研究表明，它可以提高采用不同冷冻保存方法的精子存活率。本研究评估了在传统慢速（CS）冷冻和动态玻璃化（VIT）培养基中添加RES对犬精子质量变量的影响。使用了来自30只去势成年犬的20份混合精子样本（每份样本包含三个附睾样本）。在每份混合样本中，根据RES添加情况（对照[未添加]或100µM）和冷冻保存方法（CS或VIT）设置了四种处理方式：CS-对照、CS+RES、VIT-对照和VIT+RES。CS冷冻是通过将0.25ml细管中的精子样本暴露于液氮（LN₂）蒸汽中来进行的。玻璃化是通过将30µl精子滴直接浸入LN₂中来实现的。结果初步表明，在精子运动学和膜完整性方面，VIT处理产生的结果与CS冷冻相似。与CS-对照处理的精子样本相比，CS+RES处理的精子样本在活力、存活率、顶体完整性和DNA完整性方面表现出更高的百分比（P<0.05）。此外，与CS-对照和VIT-对照处理相比，CS+RES和VIT+RES处理的精子DNA片段化百分比均较低（P<0.05）。尽管如此，RES对玻璃化后的某些运动学参数（如速度、直线性和鞭打交叉频率）以及精子头部尺寸有负面影响。总之，白藜芦醇提高了传统慢速冷冻后精子的活力、存活率和顶体完整性，并减少了DNA片段化。虽然它在玻璃化后也降低了DNA片段化，但对犬附睾精子的运动学有负面影响。
