Tvrdá Eva, Arroyo Francisca, Gosálvez Jaime
Department of Animal Physiology, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2, Nitra, 94976, Slovakia.
Unit of Genetics, Department of Biology, Universidad Autónoma de Madrid, Madrid, Spain.
Int Urol Nephrol. 2018 Aug;50(8):1381-1388. doi: 10.1007/s11255-018-1915-9. Epub 2018 Jun 20.
The purpose of the study was to evaluate the impact of seminal plasma in human ejaculates on the sperm DNA quality and DNA longevity.
Semen samples for this study were obtained from 20 donors with a normal spermiogram. Following centrifugation, the sperm pellet was resuspended either in the seminal plasma proceeding from its respective donor, or in an equal amount of PBS, adjusting the concentration to 50 × 10/ml. Each set of samples was incubated at 37 °C for 24 h and the sperm DNA damage was assessed using the chromatin-dispersion test following 0, 2, 6, and 24 h of incubation.
Sperm DNA fragmentation (SDF) did not differ between the two experimental conditions at T0; however, Kaplan-Meier estimates to test for survival showed an statistical significant increase of SDF in the seminal plasma group when compared to the PBS group following all timeframes (p 0.000). With respect to sperm DNA longevity, the most dramatic loss of sperm DNA quality occurred during the first 2 h of incubation, with the rate of SDF (rSDF) in the seminal plasma being 2.1 more intense than in PBS. Interestingly, the rSDF was found to vary significantly between individuals, which was confirmed with significant correlations in all rSDF timeframes (rSDF T0-2, p 0.049; rSDF T2-6, p 0.000; rSDF T6-24: p 0.000).
Co-incubation of semen samples in seminal plasma after ejaculation increases iatrogenic sperm damage. This effect is statistically significant after 2 h of co-incubation. Subsequently, for ART purposes seminal plasma must be quickly removed after ejaculation-liquefaction, to avoid a higher susceptibility of sperm DNA towards fragmentation.
本研究旨在评估人类射精中的精浆对精子DNA质量和DNA寿命的影响。
本研究的精液样本取自20名精子图谱正常的捐献者。离心后，将精子沉淀重悬于来自其各自捐献者的精浆中，或等量的PBS中，将浓度调整为50×10⁶/ml。每组样本在37℃孵育24小时，并在孵育0、2、6和24小时后使用染色质分散试验评估精子DNA损伤。
在T0时，两种实验条件下的精子DNA碎片化（SDF）没有差异；然而，生存检验的Kaplan-Meier估计显示，与PBS组相比，精浆组在所有时间框架后的SDF均有统计学显著增加（p<0.000）。关于精子DNA寿命，精子DNA质量最显著的损失发生在孵育的前2小时，精浆中的SDF率（rSDF）比PBS中的高2.1倍。有趣的是，发现rSDF在个体之间有显著差异，这在所有rSDF时间框架中都有显著相关性得到证实（rSDF T0-2，p<0.049；rSDF T2-6，p<0.000；rSDF T6-24：p<0.000）。
射精后精液样本与精浆共同孵育会增加医源性精子损伤。这种效应在共同孵育2小时后具有统计学显著性。随后，出于辅助生殖技术的目的，射精液化后必须迅速去除精浆，以避免精子DNA对碎片化的更高易感性。
