Biochemical and physical changes during the cryopreservation process adversely affect sperm function required for fertilization. Recently, many studies have been conducted to find effective pre-freezing treatments to limit these damages. The present study aimed to investigate the effects of pre-freezing treatment with quercetin and crocin, individually or in combination, on sperm parameters after thawing procedure. For this, semen samples from 20 normozoospermic men were collected and then each sample was divided into five equal parts: 1. fresh group 2. frozen–thawed group without addition of antioxidants 3. frozen–thawed group containing 1 mM crocin, 4. frozen–thawed group containing 50 μM quercetin, and 5. frozen–thawed group containing a combination of 1 mM crocin and 50 μM quercetin. Pre-cryopreservation and post-thaw sperm motility, morphology, viability, DNA fragmentation, reactive oxygen species [1] (ROS) levels, and mitochondrial membrane potential [2] (MMP) were investigated. Cryopreservation significantly reduced sperm quality. Both crocin and quercetin individually improved sperm progressive motility, decreased ROS levels, reduced DNA fragmentation, and marginally increased MMP, though crocin seems to be more successful in protecting sperm quality. More interestingly, the combined addition of crocin and quercetin to the sperm-freezing medium did not show positive effects on sperm quality. Crocin and quercetin may play a role in mitigating the cryopreservation-induced injury to sperm.

冷冻保存过程中的生化和物理变化会对受精所需的精子功能产生不利影响。最近，进行了许多研究来寻找有效的预冷冻处理来限制这些损害。本研究旨在探讨槲皮素和藏红花素单独或组合进行预冷冻处理对解冻后精子参数的影响。为此，收集了 20 名正常精子男性的精液样本，然后将每个样本分为五等份： 1. 新鲜组 2. 不添加抗氧化剂的冻融组 3. 含有 1 mM 藏红花素的冻融组，4. 冷冻组– 含有 50 μM 槲皮素的解冻组，以及 5. 含有 1 mM 藏红花素和 50 μM 槲皮素组合的冷冻-解冻组。研究了冷冻保存前和解冻后的精子活力、形态、活力、DNA 碎片、活性氧 [1] (ROS) 水平和线粒体膜电位 [2] (MMP)。冷冻保存显着降低了精子质量。藏红花素和槲皮素均单独改善了精子的前向运动性，降低了ROS水平，减少了DNA碎片，并略微增加了MMP，尽管藏红花素似乎在保护精子质量方面更成功。更有趣的是，在精子冷冻培养基中联合添加藏红花素和槲皮素并没有显示出对精子质量的积极影响。藏红花素和槲皮素可能在减轻冷冻保存引起的精子损伤方面发挥作用。