Thanintranon Sitthanan, Saeng-Anan Ubol, Vutyavanich Teraporn, Piromlertamorn Waraporn, Somsak Pareeya, Sanmee Usanee
Chiang Mai University Faculty of Medicine Department of Obstetrics and Gynecology Chiang Mai 50200 Thailand Division of Reproductive Medicine, Department of Obstetrics and Gynecology, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand.
Chiang Mai University Center of Medical Excellence CMEx Fertility Center Chiang Mai 50200 Thailand CMEx Fertility Center, Center of Medical Excellence, Chiang Mai University, Chiang Mai 50200, Thailand.
JBRA Assist Reprod. 2024 Dec 3;28(4):611-617. doi: 10.5935/1518-0557.20240056.
To investigate the effect of astaxanthin supplementation in cryopreservation media on post-thawed sperm motility, viability, morphology, reactive oxygen species (ROS), and DNA fragmentation in two cryopreservation techniques using vitrification and liquid nitrogen vapor freezing.
Thirty normozoospermic semen samples were used in the study. Post-prepared semen samples were divided into 1) non-cryopreserved control, 2) and 3) vitrified without (V) and with astaxanthin 0.5 µM (V+ATX), 4) and 5) frozen in liquid nitrogen vapor without (L) and with astaxanthin 0.5 µM (L+ATX).
Cryopreservation using vitrification and liquid nitrogen vapor freezing significantly decreased sperm motility and viability and increased ROS levels. However, no changes were seen in sperm morphology or DNA fragmentation. The addition of astaxanthin in cryopreservation media significantly increased post-thawed motility in both vitrification (77.6±8.9% vs. 69.0±9.5% in V+ATX and V) and vapor freezing (57.0±13.3% vs. 47.7±14.6% in L+ATX and L); it significantly increased sperm viability in vitrification (75.0±11.9% vs. 65.9±11.1% in V+ATX and V), and significantly decreased ROS level in both vitrification (4.7 (2.6-8.3) RLU/sec/106 vs. 10.6 (9.4-16.0) RLU/sec/106 in V+ATX and V) and vapor freezing (4.6 (3.3-10.5) RLU/sec/106 vs. 10.3 (7.9-18.6) RLU/ sec/106 in L+ATX and L). Astaxanthin supplementation in cryopreservation media did not affect sperm morphology or DNA fragmentation.
Astaxanthin supplementation improved post-cryopreserved sperm motility, decreased ROS levels in both vitrification and liquid nitrogen vapor freezing and improved sperm viability only in the vitrification technique.
研究在冷冻保存介质中添加虾青素对两种冷冻保存技术（玻璃化冷冻和液氮蒸汽冷冻）解冻后精子活力、存活率、形态、活性氧（ROS）及DNA片段化的影响。
本研究使用了30份正常精子精液样本。制备后的精液样本分为：1）未冷冻保存的对照组；2）和3）未添加虾青素（V）和添加0.5µM虾青素（V+ATX）的玻璃化冷冻组；4）和5）未添加虾青素（L）和添加0.5µM虾青素（L+ATX）的液氮蒸汽冷冻组。
玻璃化冷冻和液氮蒸汽冷冻显著降低了精子活力和存活率，并提高了ROS水平。然而，精子形态或DNA片段化未见变化。在冷冻保存介质中添加虾青素显著提高了玻璃化冷冻（V+ATX组为77.6±8.9%，V组为69.0±9.5%）和蒸汽冷冻（L+ATX组为57.0±13.3%，L组为47.7±14.6%）解冻后的精子活力；显著提高了玻璃化冷冻后的精子存活率（V+ATX组为75.0±11.9%，V组为65.9±11.1%），并显著降低了玻璃化冷冻（V+ATX组为4.7（2.6 - 8.3）RLU/秒/10⁶，V组为10.6（9.4 - 16.0）RLU/秒/10⁶）和蒸汽冷冻（L+ATX组为4.6（3.3 - 10.5）RLU/秒/10⁶，L组为10.3（7.9 - 18.6）RLU/秒/10⁶）中的ROS水平。在冷冻保存介质中添加虾青素不影响精子形态或DNA片段化。
添加虾青素可提高冷冻保存后精子的活力，降低玻璃化冷冻和液氮蒸汽冷冻中的ROS水平，且仅在玻璃化冷冻技术中提高精子存活率。
