Raad Georges, Mansour Joyce, Ibrahim Rim, Azoury Jessica, Azoury Joan, Mourad Youmna, Fakih Chadi, Azoury Joseph
1Azoury IVF Clinic,Mount Lebanon Hospital,Camille Chamoun Bvd,Beirut,Lebanon.
2Institut National de Pathologie (INP),Baabda,Lebanon.
Zygote. 2019 Apr;27(2):69-77. doi: 10.1017/S0967199419000030. Epub 2019 Mar 5.
SummaryDirect swim-up procedure is widely used to separate the motile competent spermatozoa from the antioxidant-rich semen. Subsequently, spermatozoa become more vulnerable to reactive oxygen species (ROS) due to their cytological characteristics. The effect of vitamin C, a highly concentrated antioxidant in the semen, on direct swim-up-enriched sperm population is not fully investigated. Therefore, the aim of the present study was to assess the effect of vitamin C on sperm functional properties during direct swim-up procedure. Semen samples were collected from 22 participants. Each semen sample was divided into several aliquots. The first portion was overlaid with sperm medium without ascorbic acid (0 µM AA). The second and third fractions were overlaid with sperm medium supplemented with 300 µM and 600 µM AA; respectively. After 1 h of incubation, basic sperm parameters, intracellular ROS levels, acrosome reaction, chromatin integrity, and glucose uptake were assessed. Swim-up without AA significantly increased the percentage of ROS(+) spermatozoa compared with the raw semen (P<0.01). Interestingly, swim-up with 300 µM AA did not increase the percentage of ROS(+) sperm compared with the raw semen. In parallel, the percentage of sperm with altered chromatin integrity was significantly lower in the 300 µM AA group compared with that in the raw semen (P<0.05). These findings suggest that supplementation of vitamin C to sperm medium could be beneficial for direct swim-up-derived spermatozoa.
直接上游法被广泛用于从富含抗氧化剂的精液中分离出具有运动能力的有功能精子。随后，由于精子的细胞学特征，它们变得更容易受到活性氧（ROS）的影响。精液中高浓度抗氧化剂维生素C对直接上游富集精子群体的影响尚未得到充分研究。因此，本研究的目的是评估维生素C在直接上游法过程中对精子功能特性的影响。从22名参与者中采集精液样本。每个精液样本被分成几个等分试样。第一部分覆盖不含抗坏血酸（0µM AA）的精子培养基。第二部分和第三部分分别覆盖添加了300µM和600µM AA的精子培养基。孵育1小时后，评估基本精子参数、细胞内ROS水平、顶体反应、染色质完整性和葡萄糖摄取。与未处理精液相比，不添加AA的上游法显著增加了ROS（+）精子的百分比（P<0.01）。有趣的是，与未处理精液相比，添加300µM AA的上游法并未增加ROS（+）精子的百分比。同时，300µM AA组中染色质完整性改变的精子百分比显著低于未处理精液组（P<0.05）。这些发现表明，在精子培养基中添加维生素C可能对直接上游法获得的精子有益。
