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猪卵母细胞氧化应激的机制及抗氧化剂的作用

Mechanisms of oxidative stress in porcine oocytes and the role of anti-oxidants.

作者信息

Whitaker B D, Knight J W

机构信息

Department of Animal and Poultry Sciences, Virginia Tech, Blacksburg, VA 24061, USA.

出版信息

Reprod Fertil Dev. 2008;20(6):694-702. doi: 10.1071/rd08037.

Abstract

The mechanisms of oxidative stress in in vitro maturing porcine oocytes and the effects of anti-oxidant supplementation of the medium in ameliorating these effects were investigated in the present study. In addition to intracellular reduced glutathione (GSH) concentrations and DNA fragmentation, the present study focused on superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase activity. The anti-oxidants used were N-acetylcysteine (NAC) and its derivative NAC-amide (NACA). The results indicate that when SOD is inhibited, supplementation of the maturarion medium with 1.5 mm NAC or NACA compensates for the decrease in SOD activity by reducing the degree of DNA fragmentation (P < 0.05). When GPx is inhibited, supplementation of the maturarion medium with 1.5 mm NAC alleviates the effects of no GPx activity, as indicated by a decrease in the degree of DNA fragmentation (P < 0.05). When the maturarion medium was supplemented with 1.5 mm NACA, intracellular GSH concentrations decreased (P < 0.05) and SOD and catalase activities increased (P < 0.05) along with the degree of DNA fragmentation. These results indicate that the mechanisms of alleviating oxidative stress in porcine oocytes are very complex and supplementing maturing oocytes with anti-oxidants may enhance enzyme activities and eliminate free radicals.

摘要

本研究调查了体外成熟猪卵母细胞中氧化应激的机制以及培养基中添加抗氧化剂对改善这些影响的作用。除了细胞内还原型谷胱甘肽(GSH)浓度和DNA片段化外,本研究还关注超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GPx)和过氧化氢酶活性。使用的抗氧化剂是N-乙酰半胱氨酸(NAC)及其衍生物NAC-酰胺(NACA)。结果表明,当SOD受到抑制时,在成熟培养基中添加1.5 mM NAC或NACA可通过降低DNA片段化程度来补偿SOD活性的下降(P < 0.05)。当GPx受到抑制时,在成熟培养基中添加1.5 mM NAC可减轻无GPx活性的影响,表现为DNA片段化程度降低(P < 0.05)。当在成熟培养基中添加1.5 mM NACA时,细胞内GSH浓度降低(P < 0.05),SOD和过氧化氢酶活性增加(P < 0.05),同时DNA片段化程度也增加。这些结果表明,减轻猪卵母细胞氧化应激的机制非常复杂,向成熟卵母细胞中添加抗氧化剂可能会增强酶活性并清除自由基。

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