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二十碳五烯酸对牛卵丘-卵母细胞复合体的体外作用。

Effect of eicosapentaenoic acid on bovine cumulus-oocyte complex in vitro.

作者信息

Nikoloff Noelia, Pascua Ana M, Anchordoquy Juan M, Anchordoquy Juan P, Sirini Matías A, Seoane Analia, Furnus Cecilia C

机构信息

IGEVET-Instituto de Genética Veterinaria "Ing. Fernando N Dulout" (UNLP-CONICET LA PLATA), Facultad de Ciencias Veterinarias UNLP, Calles 60 y 118, B1904AMA La Plata, Buenos Aires, Argentina.

出版信息

Cell Biol Int. 2017 May;41(5):505-513. doi: 10.1002/cbin.10746. Epub 2017 Mar 13.

Abstract

The aim of the present study was to investigate the effects of eicosapentaenoic acid (EPA) supplementation during in vitro maturation (IVM) of bovine oocytes. The concentrations tested in all experiments were 1 nM, 1 μM, and 1 mM EPA. The effect of EPA was evaluated on cumulus-oocyte complexes (COC) by oocyte maturation (cumulus expansion area and oocyte nuclear maturation), genotoxicity [single cell gel electrophoresis (SCGE)], and cytotoxicity (apoptosis, viability, and MTT assays) end points. The maturation parameters were affected by exposure of COC to different EPA concentrations in the IVM medium. Cumulus expansion area increased in the presence of 1 nM EPA (P < 0.05) whereas addition of 1 nM EPA (P < 0.05) decreased cumulus expansion after 24 h of IVM. Moreover, the maturation rate significantly decreased when 1 mM of EPA was assayed (P < 0.001). EPA at 1 nM induced genotoxic and cytotoxic effects on bovine cumulus cells (CC) and primary DNA lesions (P < 0.001). A significant increase in the frequency of apoptotic (P < 0.01) and necrotic (P < 0.001) cells was observed after 24 h of treatment with 1 nM, 1 μM, and 1 mM EPA. Mitochondrial activity was altered with 1 mM EPA (P < 0.001). We inferred that optimal oocyte quality was partially dependent on the presence of adequate EPA concentrations; EPA could be beneficial to improve oocyte quality in the maturation process, because low concentration tested (1 nM EPA) improved cumulus expansion.

摘要

本研究的目的是调查在牛卵母细胞体外成熟(IVM)过程中补充二十碳五烯酸(EPA)的效果。在所有实验中测试的EPA浓度为1 nM、1 μM和1 mM。通过卵母细胞成熟(卵丘扩展面积和卵母细胞核成熟)、遗传毒性[单细胞凝胶电泳(SCGE)]和细胞毒性(凋亡、活力和MTT测定)终点来评估EPA对卵丘-卵母细胞复合体(COC)的影响。成熟参数受到IVM培养基中COC暴露于不同EPA浓度的影响。在存在1 nM EPA的情况下,卵丘扩展面积增加(P < 0.05),而添加1 nM EPA(P < 0.05)在IVM 24小时后会降低卵丘扩展。此外,当检测1 mM EPA时,成熟率显著降低(P < 0.001)。1 nM的EPA对牛卵丘细胞(CC)诱导了遗传毒性和细胞毒性作用以及原发性DNA损伤(P < 0.001)。在用1 nM、1 μM和1 mM EPA处理24小时后,观察到凋亡细胞(P < 0.01)和坏死细胞(P < 0.001)的频率显著增加。1 mM EPA改变了线粒体活性(P < 0.001)。我们推断,最佳的卵母细胞质量部分取决于是否存在足够浓度的EPA;EPA可能有利于在成熟过程中提高卵母细胞质量,因为测试的低浓度(1 nM EPA)改善了卵丘扩展。

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