IGEVET - Instituto de Genética Veterinaria "Ing. Fernando N Dulout" (UNLP-CONICET LA PLATA), Facultad de Ciencias Veterinarias, Universidad Nacional de La Plata, Calle 60 y 118, B1904AMA, La Plata, Buenos Aires, Argentina.
Environ Sci Pollut Res Int. 2021 Jun;28(23):29188-29199. doi: 10.1007/s11356-021-12670-x. Epub 2021 Feb 7.
The aim of this study was to evaluate the genotoxic and cytotoxic effects of amitraz (AMZ) on the primary culture of bovine cumulus cells (CC) and oocyte nuclear maturation. Cytotoxicity was evaluated by assessing mitochondrial activity with the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Genotoxicity was estimated using the alkaline single cell gel electrophoresis (SCGE) assay. Apoptosis was detected with the Annexin V-affinity assay. The in vitro maturation test was performed in bovine oocytes. To understand AMZ action, glutathione content, superoxide dismutase enzyme activity, and lipid peroxidation were evaluated in CC. Results showed that AMZ lethal concentration (LC 50) for bovine CC was 32.55 μg/mL (MTT assay). A 25 μg/mL induced late apoptosis and necrotic cells (p < 0.05); however, DNA damage was decreased at the same concentration (SCGE assay; p < 0.05). A decrease in metaphase II was observed at 25 μg/mL, and degenerate oocytes were observed at 15 and 25 μg/mL (p < 0.05). None of the oxidative stress parameters evaluated showed significant differences. This study contributes to a better understanding of AMZ in this model, suggesting its potential cytotoxicity and impact on bovine reproduction.
本研究旨在评估双甲脒（AMZ）对牛卵丘-卵母细胞复合体（CC）原代培养物和卵母细胞核成熟的遗传毒性和细胞毒性作用。通过使用 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2H-四唑溴盐（MTT）测定法评估线粒体活性来评估细胞毒性。通过碱性单细胞凝胶电泳（SCGE）测定法估计遗传毒性。通过 Annexin V 亲和测定法检测细胞凋亡。体外成熟试验在牛卵母细胞中进行。为了了解 AMZ 的作用，评估了 CC 中的谷胱甘肽含量、超氧化物歧化酶酶活性和脂质过氧化作用。结果表明，AMZ 对牛 CC 的致死浓度（LC50）为 32.55μg/mL（MTT 测定法）。25μg/mL 诱导晚期凋亡和坏死细胞（p<0.05）；然而，在相同浓度下，DNA 损伤减少（SCGE 测定法；p<0.05）。在 25μg/mL 时观察到中期 II 减少，在 15 和 25μg/mL 时观察到退化卵母细胞（p<0.05）。评估的所有氧化应激参数均未显示出显着差异。本研究有助于更好地了解该模型中的 AMZ，表明其潜在的细胞毒性及其对牛繁殖的影响。
