Ayla Şule, Tunalı Gülden, Bilgiç Bülent E, Sofuoğlu Kenan, Özdemir A Arman, Tanrıverdi Gamze, Özdemir Semra, Soner B Cem, Öztürk Bahar, Karahüseyinoğlu Serçin, Aslan Esra Güler, Seçkin Ismail
Istanbul Medipol University, School of Medicine, Department of Histology and Embryology, Istanbul, Turkey.
Zeynep Kamil Gynecology and Maternity Training and Research Hospital, IVF-ET Unit, Istanbul, Turkey.
Acta Histochem. 2018 Feb;120(2):117-121. doi: 10.1016/j.acthis.2018.01.001. Epub 2018 Jan 8.
Sperm processing (e.g., centrifugation) used in preparation for assisted reproduction can result in excessive generation of reactive oxygen species (ROS) and potential sperm damage. The use of antioxidants during sperm processing has been shown to prevent iatrogenic sperm damage, including DNA damage. In this study, we evaluated the effect of caffeic acid phenethyl ester (CAPE) on oxidative stress mediated sperm dysfunction and DNA damage.
Semen samples were obtained to liquefy at room temperature. After centrifugation and washing protocols, spermatozoa were incubated in a single step supplemented medium with either of 10, 50 or 100 μmol/L CAPE for 2 hours at 36 °C. After incubation period, MDA levels of seminal plasma were measured. The fragmentation in sperm DNA was detected by light microscopy via use of an aniline blue assay, while ultrastructural morphology was analyzed by transmission electron microscopy.
Significant increase has been observed in percent chromatin condensation (assessed by aniline blue staining) and Malondialdehyde (Mmol/L) in oligoasthenoteratozoospermia group before the centrifugation (0.57 ± 0.15). Incubation of samples with 100 μmol/L CAPE after centrifugation resulted in a significantly lower percent chromatin condensation compared to samples incubated without CAPE (0.42 ± 0.12) (P < 0.0033). Incubation of all samples with CAPE (10 μmol/L, 50 μmol/L, 100 μmol/L.) after centrifugation resulted in a significantly lower percentage of Malondialdehyde levels.
The data suggests that preincubation of spermatozoa with the antioxidant CAPE offers protection against oxidative DNA damage in vitro.
辅助生殖准备过程中使用的精子处理方法（如离心）可导致活性氧（ROS）过度生成及潜在的精子损伤。研究表明，精子处理过程中使用抗氧化剂可预防医源性精子损伤，包括DNA损伤。在本研究中，我们评估了咖啡酸苯乙酯（CAPE）对氧化应激介导的精子功能障碍和DNA损伤的影响。
获取精液样本并在室温下液化。经过离心和洗涤程序后，将精子在36°C下于含有10、50或100μmol/L CAPE的单步补充培养基中孵育2小时。孵育期结束后，测量精浆中的丙二醛水平。通过使用苯胺蓝测定法，通过光学显微镜检测精子DNA的片段化，同时通过透射电子显微镜分析超微结构形态。
在少弱畸精子症组离心前，染色质浓缩百分比（通过苯胺蓝染色评估）和丙二醛（mmol/L）显著增加（0.57±0.15）。离心后用100μmol/L CAPE孵育样本，与未用CAPE孵育的样本相比，染色质浓缩百分比显著降低（0.42±0.12）（P<0.0033）。离心后将所有样本与CAPE（10μmol/L、50μmol/L、100μmol/L）一起孵育，丙二醛水平百分比显著降低。
数据表明，精子与抗氧化剂CAPE预孵育可在体外提供针对氧化性DNA损伤的保护作用。
