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冷冻保存后牛精子中谷胱甘肽(GSH)含量的降低:两种稀释液的比较

Decrease in glutathione (GSH) content in bovine sperm after cryopreservation: comparison between two extenders.

作者信息

Stradaioli G, Noro T, Sylla L, Monaci M

机构信息

Dipartimento di Scienze Animali, Università di Udine, via delle Scienze 208, 33100 Udine, Italy.

出版信息

Theriogenology. 2007 Apr 15;67(7):1249-55. doi: 10.1016/j.theriogenology.2007.01.009. Epub 2007 Mar 6.

DOI:10.1016/j.theriogenology.2007.01.009
PMID:17337045
Abstract

Growing evidence suggests that among the causes which deteriorate qualitative and functional characteristics of semen after freezing and thawing, there are those linked to oxidative stress and decrease detoxification of antioxidants present in the cells and seminal plasma. Overall, cryopreservation reduces the spermatozoal GSH content. In the present study we assessed the motility, capacitation status, ability to undergo acrosome reaction and levels of GSH in bovine spermatozoa after freezing and thawing, and compared the effects of the egg yolk tris-citrate and a commercial extenders (Bioxcell). Six semen samples were collected from each of five bulls and frozen and thawed in the two extenders. The level of total GSH of bovine spermatozoa detected in the raw semen ranged from 246.5 to 776.2 pmoles/mg of protein. The Bioxcell extender was superior in preserving GSH content (647.2+/-300.1 versus 223.6+/-154.0 pmoles/mL; P<0.05), motility levels (44.3+/-4.8 versus 41.8+/-4.0%; P<0.05), non-capacitated state (68.3+/-13.6 versus 61.3+/-17%; P<0.05) and a lower proportion of acrosome reacted spermatozoa post thaw (3.8+/-2.1 versus 6.3+/-3.1%; P<0.001) compared to egg yolk tris-citrate extender. These findings could be partially ascribed to the high GSH level contained in the commercial extender which seem able to alleviate oxidative damages to spermatozoa surviving freezing thawing procedures. Further researches are required in order to validate in vivo these results.

摘要

越来越多的证据表明,在冷冻和解冻后精液质量和功能特性恶化的原因中,有一些与氧化应激以及细胞和精浆中抗氧化剂解毒能力的降低有关。总体而言,冷冻保存会降低精子的谷胱甘肽(GSH)含量。在本研究中,我们评估了冷冻和解冻后牛精子的活力、获能状态、顶体反应能力以及GSH水平,并比较了卵黄三柠檬酸缓冲液和一种商业稀释液(Bioxcell)的效果。从五头公牛中每头采集六个精液样本,并在两种稀释液中进行冷冻和解冻。在新鲜精液中检测到的牛精子总GSH水平为246.5至776.2皮摩尔/毫克蛋白质。与卵黄三柠檬酸缓冲液稀释液相比,Bioxcell稀释液在保存GSH含量(647.2±300.1对223.6±154.0皮摩尔/毫升;P<0.05)、活力水平(44.3±4.8对41.8±4.0%;P<0.05)、未获能状态(68.3±13.6对61.3±17%;P<0.05)以及解冻后顶体反应精子的比例较低(3.8±2.1对6.3±3.1%;P<0.001)方面更具优势。这些发现可能部分归因于商业稀释液中所含的高GSH水平,它似乎能够减轻冷冻解冻过程中存活精子的氧化损伤。为了在体内验证这些结果,还需要进一步的研究。

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