成熟液中添加白藜芦醇对猪卵母细胞体外发育和抗冻能力的影响

doi:10.11924/j.issn.1000-6850.casb17070057

中国农学通报 ›› 2018, Vol. 34 ›› Issue (28): 137-142.doi: 10.11924/j.issn.1000-6850.casb17070057

所属专题：生物技术；畜牧兽医

成熟液中添加白藜芦醇对猪卵母细胞体外发育和抗冻能力的影响

戴建军,吴彩凤,张树山,葛雷,陈亚宁,张德福

上海市农业科学院畜牧兽医研究所,上海市农业科学院畜牧兽医研究所,上海市农业科学院畜牧兽医研究所,上海市农业科学院畜牧兽医研究所,上海市农业科学院畜牧兽医研究所,上海市农业科学院畜牧兽医研究所

收稿日期:2017-07-12 修回日期:2018-09-06 接受日期:2017-12-20 出版日期:2018-10-11 发布日期:2018-10-11
通讯作者: 张德福
基金资助:
国家自然科学基金“猪胚胎玻璃化冷冻保存对其细胞凋亡模式和基因表达的影响”(31372315)；上海市自然科学基金“基于转录组学与蛋白质组学的猪精子耐冻性影响机制研究”(15ZR1430100)；上海市农业科学院攀高计划(PG171)。

RES Addition in IVM Medium Affects Development and Anti-freezing Ability of Porcine MII Stage Oocytes in Vitro

Received:2017-07-12 Revised:2018-09-06 Accepted:2017-12-20 Online:2018-10-11 Published:2018-10-11

摘要/Abstract

摘要： 为探究猪卵母细胞成熟培养液中添加白藜芦醇(Resveratrol,RES)对其体外发育与抗冻能力的影响。本试验通过添加不同浓度的RES,观察其对卵母细胞体外发育和相关基因的表达的影响；通过OPS(Open Pulled Straw)玻璃化冷冻和解冻,观察RES对卵母细胞冷冻前后线粒体功能、凋亡和发育能力的影响。结果表明：(1)成熟培养液中添加2 μmol/L RES有利于提高猪卵母细胞的体外成熟和发育能力,提高RES浓度至10 μmol/L时则表现为细胞毒性作用；(2)添加2 μmol/L RES可显著提高卵母细胞Sirt1,MATER,SOD1和Bcl2基因的表达量,并显著降低c-mos,MAPK和Mfn2的表达；当RES添加量增至10 μmol/L时,卵母细胞凋亡相关基因的表达呈现出促凋亡状态；(3)添加RES后,成熟卵母细胞在冷冻前后的ROS水平均要低于未添加组,ATP含量和线粒体膜电位值则高于未添加组；(4)成熟液中添加RES后,卵母细胞冻后FDA染色存活率和孤雌激活卵裂率均高于未添加组,,早期凋亡率则低于未添加组,其中FDA染色存活率差异显著。综上所述,2 μmol/L的RES在猪卵母细胞成熟液中添加,可通过改善其发育和凋亡相关基因的表达,提高线粒体功能,达到提高卵母细胞发育和抗冻能力的目的。

Abstract: The purpose of this study was to clarify the effect of RES addition in IVM medium on in vitro development and anti-freezing ability of porcine oocytes. Different concentrations of RES were added in IVM media, the in vitro developmental ability and the expression levels of related genes were analyzed. After open pulled straw (OPS) vitrification and thawing, the mitochondrial membrane potential (ΔΨm), early stage apoptotic level and parthenogenetic developmental ability of porcine oocytes were analyzed. The results were as follows. (1) 2 μmol/L RES addition could improve the IVM rate and parthenogenetic cleavage rate, a high concentration of RES addition (10 μmol/L) showed a cytotoxic effect on the developmental ability; (2) 2 μmol/L RES addition in IVM medium significantly improved the expression levels of Sirt1, MATER, SOD1 and Bcl2, and decreased the expression levels of c-mos, MAPK and Mfn2; when the RES was 10 μmol/L, the expression of apoptotic related genes presented proapoptotic status; (3) ROS levels in fresh or vitrified oocytes of RES addition groups were lower than that of control group, ATP contents and ΔΨm values were higher than those of control group without RES addition; (4) RES addition in IVM medium increased the FDA survival and parthenogenetic cleavage rates, and decreased the early stage apoptotic rate than those of control group without RES addition. In conclusion, 2 μmol/L RES addition in porcine IVM medium can lead to better gene expression levels, increase mitochondrial function, then improve the development and anti-freezing ability of porcine oocytes

戴建军,吴彩凤,张树山,葛雷,陈亚宁,张德福. 成熟液中添加白藜芦醇对猪卵母细胞体外发育和抗冻能力的影响[J]. 中国农学通报, 2018, 34(28): 137-142.

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成熟液中添加白藜芦醇对猪卵母细胞体外发育和抗冻能力的影响

RES Addition in IVM Medium Affects Development and Anti-freezing Ability of Porcine MII Stage Oocytes in Vitro

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