Li Shi-Jia, Su Wei-Dong, Qiu Li-Jun, Wang Xiong, Liu Juan
Department of Clinical Laboratory, Tianjin 4th Central Hospital, Tianjin 310140, China.
Center of Reproductive Medicine, Yantai Yuhuangding Hospital, Yantai, Shandong 264000, China.
Zhonghua Nan Ke Xue. 2018 Jun;24(6):499-503.
To investigate the effects of resveratrol in the cryopreservation medium on the quality and function of post-thaw sperm.
Semen samples were obtained from 50 normozoospermic and 50 oligoasthenozoospermic men, liquefied and then cryopreserved in the glycerol-egg yolk-citrate (GEYC) medium with or without 30 μmol/L resveratrol. Sperm motility, viability and acrosome reaction (AR) were examined before and after thawing. Sperm lipid peroxidation and the level of reactive oxygen species (ROS) were measured using commercial malondialdehyde (MDA) and the ROS assay kit. Sperm mitochondrial membrane potential (MMP) and DNA damage were determined by Rhodamine 123 staining and TUNEL.
The percentage of progressively motile sperm (PMS), total sperm motility, sperm viability, MMP and AR were significantly decreased (P <0.05) while the levels of sperm ROS, MDA and DNA fragmentation index (DFI) remarkably increased in both the normozoospermia and oligoasthenozoospermia groups after cryopreservation as compared with those in the fresh ejaculate (P <0.05). In comparison with the non-resveratrol control, the post-thaw sperm cryopreserved with 30 μmol/L resveratrol showed markedly higher PMS ([32.7 ± 4.8] vs [43.1 ± 6.3] %, P <0.05), total motility ([44.8 ± 6.9] vs [56.9 ± 7.4] %, P <0.05), viability ([52.3 ± 6.1] vs [67.5 ± 5.6] %, P <0.05), MMP ([56.5 ± 7.0] vs [63.4 ± 7.5] %, P <0.05) and AR ([16.6 ± 3.8] vs [26.3 ± 4.7] %, P <0.05) but lower ROS, MDA and DFI (all P <0.05) in the normozoospermia group, and so did the post-thaw sperm in the oligoasthenozoospermia group, with a particularly lower DFI ([28.5 ± 4.8] vs [36.3 ± 5.7]%, P <0.01).
Resveratrol in the cryopreservation medium can improve the quality and function of post-thaw human sperm by reducing cryopreservation-induced sperm injury and the level of ROS.
探讨白藜芦醇添加到冷冻保存培养基中对解冻后精子质量和功能的影响。
收集50例正常精子症男性和50例少弱精子症男性的精液样本,液化后分别在含或不含30 μmol/L白藜芦醇的甘油 - 蛋黄 - 柠檬酸盐(GEYC)培养基中冷冻保存。解冻前后检测精子活力、存活率及顶体反应(AR)。使用商业化的丙二醛(MDA)和活性氧(ROS)检测试剂盒测定精子脂质过氧化水平和ROS水平。通过罗丹明123染色和TUNEL法检测精子线粒体膜电位(MMP)和DNA损伤情况。
与新鲜精液相比,正常精子症组和少弱精子症组冷冻保存后,进行性运动精子(PMS)百分比、总精子活力、精子存活率、MMP和AR均显著降低(P <0.05),而精子ROS、MDA水平及DNA碎片指数(DFI)显著升高(P <0.05)。与未添加白藜芦醇的对照组相比,添加30 μmol/L白藜芦醇冷冻保存的解冻后精子,正常精子症组的PMS([32.7 ± 4.8]% 对 [43.1 ± 6.3]%,P <0.05)、总活力([44.8 ± 6.9]% 对 [56.9 ± 7.4]%,P <0.05)、存活率([52.3 ± 6.1]% 对 [67.5 ± 5.6]%,P <0.05)、MMP([56.5 ± 7.0]% 对 [63.4 ± 7.5]%,P <0.05)和AR([16.6 ± 3.8]% 对 [26.3 ± 4.7]%,P <0.05)显著更高,但ROS、MDA和DFI更低(均P <0.05);少弱精子症组解冻后精子也有类似情况,且DFI更低([28.5 ± 4.8]% 对 [36.3 ± 5.7]%,P <0.01)。
冷冻保存培养基中的白藜芦醇可通过减少冷冻保存诱导的精子损伤和ROS水平来提高解冻后人精子的质量和功能。