IGEVET-Instituto de Genética Veterinaria "Prof. Fernando N. Dulout" (UNLP-CONICET LA PLATA), Facultad de Ciencias Veterinarias, Universidad Nacional de La Plata, calle 60 y 118 s/n, 1900, La Plata, Buenos Aires, Argentina.
Cátedra de Fisiología, Laboratorio de Nutrición Mineral, Facultad de Ciencias Veterinarias, Universidad Nacional de La Plata, calle 60 y 118 s/n, 1900, La Plata, Buenos Aires, Argentina.
Environ Sci Pollut Res Int. 2017 Aug;24(24):20041-20049. doi: 10.1007/s11356-017-9683-0. Epub 2017 Jul 11.
The aim of this study was to investigate the cytotoxic and genotoxic effects of high copper (Cu) concentrations on bovine cumulus cells (CCs) cultured in vitro. We evaluated the effect of 0, 120, 240, and 360 μg/dL Cu added to in vitro maturation (IVM) medium on CC viability assessed by the trypan blue (TB)-fluorescein diacetate (FDA) and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assays, apoptosis, and DNA damage. Differences in cell viability assessed by TB-FDA were not significant among CC treated with 0, 120, 240, and 360 μg/dL Cu. However, mitochondrial activity assessed by MTT was lower in CC cultured with 120, 240, and 360 μg/dL Cu as compared with the control (p < 0.01). Percentages of apoptotic cells were higher when CCs were treated with 120, 240, and 360 μg/dL Cu (p < 0.05) due to higher frequencies of late apoptotic cells (p < 0.05). The frequency of live cells diminished in a dose-dependent manner when Cu was added to the culture medium. Whereas genetic damage index (GDI) increased significantly in CC cultured in the presence of 240 and 360 μg/dL Cu (p ˂ 0.05), DNA damage increased at all Cu concentrations tested (p ˂ 0.05). These results indicate that Cu induces cytotoxic and genotoxic effects in bovine CC.
本研究旨在探讨高浓度铜（Cu）对体外培养的牛卵丘细胞（CC）的细胞毒性和遗传毒性作用。我们评估了向体外成熟（IVM）培养基中添加 0、120、240 和 360μg/dL Cu 对 CC 活力的影响，活力通过台盼蓝（TB）-荧光素二乙酸酯（FDA）和 3-（4,5-二甲基-2-噻唑基）-2,5-二苯基-2H-四唑溴盐（MTT）测定，细胞凋亡和 DNA 损伤。用 TB-FDA 评估的 CC 活力在 0、120、240 和 360μg/dL Cu 处理的 CC 之间没有显著差异。然而，与对照组相比，用 120、240 和 360μg/dL Cu 培养的 CC 的线粒体活性（MTT 测定）较低（p<0.01）。当用 120、240 和 360μg/dL Cu 处理 CC 时，凋亡细胞的比例更高（p<0.05），这是由于晚期凋亡细胞的频率更高（p<0.05）。当 Cu 添加到培养基中时，活细胞的比例呈剂量依赖性下降。虽然在存在 240 和 360μg/dL Cu 的情况下 CC 的遗传损伤指数（GDI）显著增加（p<0.05），但在所有测试的 Cu 浓度下 DNA 损伤均增加（p<0.05）。这些结果表明 Cu 对牛 CC 具有细胞毒性和遗传毒性作用。
