Thuwanut P, Chatdarong K, Techakumphu M, Axnér E
Department of Clinical Sciences, P.O. Box 7054, Faculty of Veterinary Medicine and Animal Science, Swedish University of Agricultural Sciences (SLU), Uppsala, Sweden.
Theriogenology. 2008 Jul 15;70(2):233-40. doi: 10.1016/j.theriogenology.2008.04.005. Epub 2008 May 22.
Antioxidants partially ameliorated the negative effects of reactive oxygen species (ROS) produced during cryopreservation. The objective of the present study was to investigate the effect of cysteine and a water-soluble vitamin E analogue on the quality of frozen-thawed epididymal cat spermatozoa. Epididymal spermatozoa were collected from eight male cats and divided into three aliquots; these were resuspended with a tris egg yolk extender I (EE-I), or the same extender supplemented with 5mM dl-cysteine (EE-C) or with 5mM of a water-soluble vitamin E analogue (EE-Ve). Prior to the freezing step, sperm suspensions were added to the extender with Equex STM paste (EE-II). Sperm motility, progressive motility, membrane integrity, and acrosome status were evaluated at collection, after cooling, and at 0, 2, 4, and 6h post-thaw. Sperm DNA integrity was evaluated at 0 and 6h post-thaw. Relative to the control group, supplementation with vitamin E improved (P<0.05) post-thaw motility (69.4+/-5.6%), progressive motility (3.9+/-0.3), and membrane integrity (65.1+/-8.1%) immediately after thawing, whereas cysteine supplementation improved (P<0.05) post-thaw motility after 2h of incubation (53.8+/-12.2%) and DNA integrity after 6h (84.1+/-4.4%). However, neither antioxidant significantly increased the acrosome integrity of frozen-thawed spermatozoa. In conclusion, cysteine or vitamin E supplementation of tris egg yolk extender improved motility, progressive motility and integrity of the sperm membrane and DNA of frozen-thawed epididymal cat spermatozoa.
抗氧化剂可部分改善冷冻保存过程中产生的活性氧(ROS)的负面影响。本研究的目的是探讨半胱氨酸和一种水溶性维生素E类似物对冻融后附睾猫精子质量的影响。从八只雄性猫收集附睾精子并分成三份;将这些精子用Tris蛋黄稀释液I(EE-I)重悬,或用添加了5mM dl-半胱氨酸(EE-C)或5mM水溶性维生素E类似物的相同稀释液(EE-Ve)重悬。在冷冻步骤之前,将精子悬浮液加入含有Equex STM糊剂的稀释液(EE-II)中。在收集时、冷却后以及解冻后0、2、4和6小时评估精子活力、前向运动能力、膜完整性和顶体状态。在解冻后0和6小时评估精子DNA完整性。相对于对照组,补充维生素E可改善(P<0.05)解冻后立即的精子活力(69.4±5.6%)、前向运动能力(3.9±0.3)和膜完整性(65.1±8.1%),而补充半胱氨酸可改善(P<0.05)孵育2小时后的解冻后精子活力(53.8±12.2%)和6小时后的DNA完整性(84.1±4.4%)。然而,两种抗氧化剂均未显著提高冻融后精子的顶体完整性。总之,在Tris蛋黄稀释液中补充半胱氨酸或维生素E可改善冻融后附睾猫精子的活力、前向运动能力以及精子膜和DNA的完整性。