| 脱氧雪腐烯醇(DON)对牛卵母细胞体外成熟及发育能力的影响 |
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| 引用本文： | 孙欢,孙克佳,姜晓龙,刘爱菊,马晓菲,韩红叶,姚大为,马毅,田树军.脱氧雪腐烯醇(DON)对牛卵母细胞体外成熟及发育能力的影响[J].畜牧兽医学报,2021,52(9):2475-2483. |
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| 作者姓名： | 孙欢 孙克佳 姜晓龙 刘爱菊 马晓菲 韩红叶 姚大为 马毅 田树军 |
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| 作者单位： | 1. 河北农业大学动物科技学院, 保定 071001;2. 天津市畜牧兽医研究所, 天津 300381;3. 河北省牛羊胚胎技术创新中心, 保定 071001 |
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| 基金项目： | 河北省重点研发计划（19226635D）；天津市奶牛产业岗位技术体系（ITTCRS2017000） |
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| 摘 要： | 本试验旨在探究脱氧雪腐烯醇（DON）对牛卵母细胞体外成熟发育的影响及其作用机制。将牛卵丘卵母细胞复合体分别在含DON浓度为0、50、250、500、1 000 ng·mL-1的体外成熟培养液中进行体外成熟，检测卵丘细胞扩展程度及第一极体排出率，构建DON毒性模型。然后，借助此模型研究DON对卵母细胞的线粒体分布及后续受精卵裂率、囊胚发育率的影响，探究DON对牛卵母细胞体外成熟及后续发育能力的影响；通过检测体外成熟卵母细胞内的氧化相关因子（ROS、GSH）水平和抗氧化基因CAT、GPx4的mRNA表达量，揭示DON影响牛卵母细胞体外发育能力的分子机制。结果表明，250、500 ng·mL-1的DON显著抑制卵母细胞的第一极体排出（P<0.05），1 000 ng·mL-1的DON极显著抑制第一极体排出（P<0.01）； 250 ng·mL-1的DON显著抑制卵丘卵母细胞扩展（P<0.05），500、1 000 ng·mL-1的DON极显著抑制卵丘细胞扩展（P<0.01）；后续试验选取DON浓度为500 ng·mL-1作为毒性模型（DON组），与不含DON组（对照组）进行比较研究，结果发现，对照组与DON组的线粒体均匀分布比例（60.2%vs.40.0%）存在显著差异（P<0.05）；试验组较对照组的受精卵裂率（33.6±3.6%vs.（67.7±2.6）%）及早期囊胚率（（0.0±0.0）%vs.（18.3±2.2）%）均显著降低（P<0.05）；试验组较对照组，卵母细胞内ROS水平（1.6 vs.1.0）显著升高（P<0.05），GSH相对水平（0.4 vs.1.0）显著降低（P<0.05），抗氧化基因CAT与GPx4的mRNA相对表达量（0.0 vs.1.0；0.6 vs.1.0）显著降低（P<0.05）。以上研究表明，DON对卵母细胞的体外成熟及早期胚胎发育具有抑制作用，其作用机制与DON破坏牛卵母细胞内抗氧化系统平衡相关。
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| 关 键 词： | 脱氧雪腐烯醇 牛 卵母细胞 体外成熟 |
| 收稿时间： | 2021-03-01 |
Effect of Deoxynivalenol(DON) on Maturation and Development of Bovine Oocytes in vitro |
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| SUN Huan,SUN Kejia,JIANG Xiaolong,LIU Aiju,MA Xiaofei,HAN Hongye,YAO Dawei,MA Yi,TIAN Shujun.Effect of Deoxynivalenol(DON) on Maturation and Development of Bovine Oocytes in vitro[J].Acta Veterinaria et Zootechnica Sinica,2021,52(9):2475-2483. |
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| Authors: | SUN Huan SUN Kejia JIANG Xiaolong LIU Aiju MA Xiaofei HAN Hongye YAO Dawei MA Yi TIAN Shujun |
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| Institution: | 1. College of Animal Science and Technology, Hebei Agricultural University, Baoding 071001, China;2. Tianjin Animal Science and Veterinary Research Institute, Tianjin 300381, China;3. Hebei Technology Innovation Center of Cattle and Sheep Embryo, Baoding 071001, China |
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| Abstract: | This study aimed to investigate the effect and mechanism of deoxynivalenol (DON) on the maturation and development of bovine oocytes in vitro. Bovine oocytes were matured in vitro in the mature solution containing different concentrations of DON (0,50,250,500,1 000 ng·mL-1), respectively. The expansion degree of cumulus cell and first polar body discharge rate were detected to construct DON toxicity model. Mitochondrial distribution of oocytes, fertile cleavage rate, blastocyst development rate were determined to evaluate the effect of DON on bovine cocytes in vitro maturation and development. And the oxidative stress,where the level of the ROS and GSH and the mRNA expression of CAT、GPx4 were detected to reveal the mechanism of DON action. It was found that the first polar body excretion of the oocytes was reduced by 250 and 500 ng·mL-1 (P<0.05) and 1 000 ng·mL-1 of DON(P<0.01).The expansion degree of cumulus cells was significant inhibited by 250 ng·mL-1 of DON (P<0.05), extremely significant inhibited by 500 and 1 000 ng·mL-1 of DON (P<0.01). In subsequent experiments, 500 ng·mL-1 of DON was selected as the toxicity model (DON group) to compare with the group without DON (control group). The results showed that the mitochondrial uniform distribution was significanly different between the control group and DON group (60.2% vs. 40.0%, P<0.05). While fertile cleavage rate ((33.6±3.6)% vs. (67.7±2.6)%, P<0.05) and blastocyst rate ((0.0±0.0)% vs.(18.3±2.2)%, P<0.05) were significantly reduced in DON group compared with the control group. The relative level of ROS in the DON group was higher than that in the control group (1.6 vs.1.0, P<0.05), however, the relative level of GSH in the DON group was lower than that in the control group (0.4 vs.1.0, P<0.05), and the relative mRNA expression level of antioxidant related genes CAT and GPX4 in the DON group were lower than those in the control group (0.3 vs.1.0; 0.6 vs. 1.0, P<0.05). The results indicates that DON inhibits the maturation in vitro and early embryo development of bovine oocyte, and the mechanism is related to DON disturbing the balance of antioxidant system of bovine oocytes. |
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| Keywords: | deoxynivalenol bovine oocytes in vitro maturation |
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