Tareq K M A, Akter Quzi Sharmin, Takagi Yuji, Hamano Koh-Ichi, Sawada Tomio, Tsujii Hirotada
Laboratory of Animal Biotechnology, Faculty of Agriculture, Interdisciplinary Graduate School of Science and Technology Shinshu University 8304 Minamiminowa-mura 399-4598 Nagano Japan.
The Sawada Women's Clinic Nagoya Reproduction Center 464-0819 Nagaya Japan.
Reprod Med Biol. 2009 Nov 28;9(2):73-81. doi: 10.1007/s12522-009-0041-x. eCollection 2010 Jun.
Selenium (Se) and vitamin E (Vit-E), as an integral part of antioxidant systems, play an important role in the motility and acrosome reaction (AR) of mammalian spermatozoa. The purpose of this study was to investigate the effect of Se and Vit-E on motility, viability, AR and accumulation of ammonia in the culture medium during different incubation periods in porcine sperm.
Sperm samples were washed, swum-up and incubated at 37°C for 1 and 3 h in Sp-TALP medium supplemented with sodium selenite (SS), seleon l-methionine (SeMet) and Vit-E in the presence or absence of ammonia. Sperm motility was determined on the basis of movement quality examined by phase microscopy. Viability and AR of spermatozoa were assessed by Hoechst 33258 and chlortetracycline (CTC) staining technique, and accumulation of ammonia was measured by the indophenol method. The incorporation of C(U)-glucose was assessed with a liquid scintillation counter.
In experiment 1, the sperm motility, viability, AR and incorporation of C(U)-glucose increased significantly ( < 0.05) in SS, SeMet and Vit-E (5, 5 μg/l and 1.0 mM, respectively) compared with the control. In experiment 2, treatment of the sperm with SeMet and SeMet + Vit-E in the presence of 300 μM ammonia also resulted in a significant increase ( < 0.05) in the rate of motility, viability, AR and incorporation of C(U)-glucose. In contrast, the accumulation of ammonia was reduced by SeMet and SeMet + Vit-E compared with the other treatments.
These findings indicate that SeMet and SeMet + Vit-E may play an important role in reducing the accumulation of ammonia and subsequently in increasing the rate of AR and the utilization of glucose in porcine spermatozoa.
硒（Se）和维生素E（Vit-E）作为抗氧化系统的一个组成部分，在哺乳动物精子的活力和顶体反应（AR）中发挥重要作用。本研究的目的是探讨Se和Vit-E对猪精子在不同孵育期的活力、生存力、AR以及培养基中氨积累的影响。
将精子样本洗涤、上浮，并在补充有亚硒酸钠（SS）、硒代L-蛋氨酸（SeMet）和Vit-E的Sp-TALP培养基中于37°C孵育1小时和3小时，有无氨存在。根据相差显微镜检查的运动质量来测定精子活力。通过Hoechst 33258和金霉素（CTC）染色技术评估精子的生存力和AR，并通过靛酚法测量氨的积累。用液体闪烁计数器评估C（U）-葡萄糖的掺入。
在实验1中，与对照组相比，SS、SeMet和Vit-E（分别为5、5μg/l和1.0mM）组的精子活力、生存力、AR和C（U）-葡萄糖掺入显著增加（<0.05）。在实验2中，在300μM氨存在下用SeMet和SeMet + Vit-E处理精子也导致活力、生存力、AR和C（U）-葡萄糖掺入率显著增加（<0.05）。相比之下，与其他处理相比，SeMet和SeMet + Vit-E减少了氨的积累。
这些发现表明，SeMet和SeMet + Vit-E可能在减少猪精子中氨的积累以及随后提高AR率和葡萄糖利用方面发挥重要作用。
