白藜芦醇对绵羊卵母细胞体外受精的影响

doi:10.16431/j.cnki.1671-7236.2020.12.023

摘要/Abstract

摘要： 本试验旨在研究不同浓度（0、0.5、2.0、5.0 μmol/L）白藜芦醇（resveratrol，RES）对绵羊卵母细胞体外受精（in vitro fertilization，IVF）、卵母细胞抗氧化能力及卵丘细胞分泌类固醇激素的影响。绵羊卵母细胞在含不同浓度RES的体外成熟（in vitro maturation，IVM）液中培养24 h以后进行体外受精，并收集IVM液，测定超氧化物歧化酶（superoxide dismutase，SOD）和谷胱甘肽过氧化物酶（glutathione peroxidase，GSH-Px）的活性及脂质过氧化产物丙二醛（monochrome display adapter，MDA）的含量；用酶联免疫法测定雌二醇（estradiol，E₂）和孕酮（progesterone，P₄）的浓度。研究结果表明，与对照组相比，在IVM液中添加0.5 μmol/L RES显著提高卵裂率（P<0.05），但对受精率和囊胚率没有显著影响（P>0.05）；5.0 μmol/L RES显著降低受精率、卵裂率和囊胚率（P<0.05），对胚胎发育有抑制作用；在IVM和体外培养（in vitro culture，IVC）液中分别添加0.5 μmol/L RES均显著提高受精率、卵裂率和囊胚率（P<0.05）。与对照组相比，在IVM液中添加RES对卵丘细胞分泌E₂有一定的抑制作用，5.0 μmol/L RES显著降低E₂浓度（P<0.05）；0.5 μmol/L RES显著提高卵丘细胞P₄分泌量（P<0.05）。0.5和2.0 μmol/L RES增加SOD和GSH-Px等酶的活性，但与对照组相比无显著差异（P>0.05），却显著降低MDA含量（P<0.05）；而5.0 μmol/L RES显著降低抗氧化酶活性并增加MDA含量（P<0.05）。综上所述，在IVM和IVC液中同时添加0.5 μmol/L RES，通过增强卵母细胞抗氧化能力和P₄的浓度，并降低MDA含量，从而提高胚胎卵裂率和囊胚率。

关键词: 绵羊卵母细胞; 白藜芦醇; 体外受精; 抗氧化酶; 类固醇激素

Abstract: The purpose of this study was to investigate the effects of resveratrol (RES) at different concentrations (0,0.5,2.0,5.0 μmol/L) on in vitro fertilization (IVF) and antioxidant capacity of ovine oocytes and the secretion of steroid hormones by cumulus cells.Sheep oocytes were fertilized in vitro after maturated in different concentrations of RES for 24 h,and the in vitro maturation (IVM) medium was collected for detecting the enzyme activity of superoxide dismutase (SOD),glutathione peroxidase (GSH-Px) and the content of monochrome display adapter (MDA).The ELISA method was used to detect the concentration of estradiol (E₂) and progesterone (P₄).The results showed that when compared to the control group,adding 0.5 μmol/L RES to the IVM medium significantly increased the cleavage rate (P<0.05),but had no significant effect on the fertilization rate and blastocyst rate (P>0.05);5.0 μmol/L RES significantly reduced the fertilization rate,cleavage rate and blastocyst rate (P<0.05),which had an inhibitory effect on embryonic development.Adding 0.5 μmol/L RES to IVM and IVC (in vitro culture,IVC) medium significantly increased the fertilization rate,cleavage rate and blastocyst rate (P<0.05).Compared to the control group,the addition of RES to IVM solution had a certain inhibitory effect on the secretion of E₂ by cumulus cells,5.0 μmol/L RES significantly reduced E₂ concentration (P<0.05);0.5 μmol/L RES significantly increased the P₄ secretion of cumulus cells (P<0.05).0.5 and 2.0 μmol/L RES increased the activity of SOD,GSH-Px and other enzymes,but there was no significant difference when compared with the control group (P>0.05),but significantly reduced MDA content (P<0.05),while 5.0 μmol/L RES significantly reduced the activity of antioxidant enzymes and increased the content of MDA (P>0.05).In conclusion,0.5 μmol/L RES simultaneously added to IVM and IVC medium could enhance the antioxidant capacity of oocytes and concentration of P₄,and reduced the content of MDA,thus improved the cleavage rate and blastocyst rate.

Key words: sheep oocyte; resveratrol; in vitro fertilization; antioxidase; steroid hormone

中图分类号:

S823.34

努日比娅姆·麦麦提托合提, 赵茜, 艾日夏提·地里夏提, 张博, 阿布力孜·吾斯曼. 白藜芦醇对绵羊卵母细胞体外受精的影响[J]. 中国畜牧兽医, 2020, 47(12): 4000-4007.

NURIBIYAMU·Maimaitituoheti, ZHAO Xi, AIRIXIATI·Dilixiati, ZHANG Bo, ABULIZ·Wusiman. Effects of Resveratrol on in vitro Fertilization of Sheep Oocytes[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(12): 4000-4007.

参考文献

[1] RICHARD F J.Effects of follicular cells on oocyte maturation Ⅱ:Theca cell inhibition of bovine oocyte maturation in vitro[J].Biology of Reproduction,1996,54(1):22-28.
[2] 欧阳小娥,胡蓉.氧化应激对卵母细胞发育及体外受精-胚胎移植结局的影响[J].国际生殖健康/计划生育杂志,2016,35(4):299-302. OUYANG X E,HU R.Effects of oxidative stress on oocyte development and outcome of in vitro fertiliza-tionembryo transfer[J].Journal of International Reproductive Health/Family Planning,2016,35(4):299-302.(in Chinese)
[3] ZHIGANG S,ZHINA L I,MIN L I,et al.Research progress on pharmacological effects of resveratrol[J].Journal of Huaihai Institute of Technology (Natural Science Edition),2017,2(26):40-43.
[4] ZHANG L,WU S,XIONG Z Y.Biological effects of resveratrol and the prospect of its application in sports medicine[J].Journal of Physical Education,2005,3(12):57-60.
[5] 努日比娅姆·麦麦提托合提,张博,赵茜,等.白藜芦醇对绵羊卵母细胞体外成熟的影响[J].中国畜牧杂志,2019,55(11):105-108. MAIMAITITUOHETI N,ZHANG B,ZHAO X,et al.Effects of resveratrol on in vitro maturation of sheep oocytes[J]. Chinese Journal of Animal Science,2019,55(11):105-108.(in Chinese)
[6] MUKHERJEE A,MALIK H,SAHA A P,et al.Resveratrol treatment during goat oocytes maturation enhances developmental competence of parthenoge-netic and hand-made cloned blastocysts by modulating intracellular glutathione level and embryonic gene expression[J]. Journal of Assisted Reproduction Genetics,2014,31(2):229-239.
[7] ZABIHI A,SHABAN H,HAJARIAN H,et al.Resveratrol addition to in vitro maturation and in vitro culture media enhances developmental competence of sheep embryos[J].Domestic Animal Endocrinology,2019,28:25-31
[8] WANG F,TIAN X,ZHANG L,HE C,et al.Beneficial effect of resveratrol on bovine oocyte maturation and subsequent embryonic development after in vitro fertilization[J].Fertility and Sterility,2013,101(2):577-586.
[9] 潘鹏,谢红月,蒋钦杨,等.白藜芦醇抗氧化功能的研究进展[J].饲料工业,2020,41(2):8-14. PAN P,XIE H Y,JIANG Q Y,et al.Research progress on antioxidant function of resveratrol[J].Feed Industry,2020,41(2):8-14.(in Chinese)
[10] LI L Z,DE B H U,LI J C,et al.Effect of co-culture on the reactive oxygen species content in mouse embryos[J].Journal of Northeast Agricultural University,2013,3(44):11-16.
[11] HUANG F J,CHIN T Y,CHAN W H.Resveratrol protects against methylglyoxal-induced apoptosis and disruption of embryonic development in mouse blastocysts[J].Environmental Toxicology,2012,28(8):431-441.
[12] LEE K,WANG C,CHAILLE J M,et al.Effect of resveratrol on the development of porcine embryos produced in vitro[J].Journal of Reproduction and Development,2010,56(3):330-335.
[13] SALZANO A,ALBERO G,ZULLO G,et al.Effect of resveratrol supplementation during culture on the quality and cryotolerance of bovine in vitro produced embryos[J]. Animal Reproduction Science,2014,151(4):91-96.
[14] LI Y,WANG J,ZHANG Z,et al.Resveratrol compares with melatonin in improving in vitro porcine oocyte maturation under heat stress[J].Journal of Animal Science and Biotechnology,2016,7:33.
[15] 陈培芳.白藜芦醇对小鼠早期胚胎发育及合子基因组激活的影响[D].南宁:广西大学,2016. CHEN P F.Effects of resveratrol on early embryonic development and zygotic gene activation in mice[D].Nanning:Guangxi University,2016.(in Chinese)
[16] CASTRO F C,SCHEFER L,SCHWARZ K L,et al.Effects of melatonin on in vitro maturation of bovine oocytes and gene expression in cumulus cells:Preliminary results[J]. Reproduction,Fertility and Development, 2018,30(1) 226-227.
[17] TORRES V,HAMDI M,MILLÁN D L,et al.Resveratrol-cyclodextrin complex affects the expression of genes associated with lipid metabolism in bovine in vitro produced embryos[J].Reproduction in Domestic Animals, 2018,53(4):850-858.
[18] ORTEGA I,WONG D H,VILLANUEVA J A,et al.Effects of resveratrol on growth and function of rat ovarian granulosa cells[J]. Fertility & Sterility,2012,98(6):1563-1573.
[19] SHIN S,JEON J H,PARK D,et al.Trans-resveratrol relaxes the corpus cavernosum ex vivo and enhances testosterone levels and sperm quality in vivo[J].Archives of Pharmacal Research,2008,31(1):83-87.
[20] BASLY J P,FRANÇOISE M F,BAIL J C,et al.Estrogenic/antiestrogenic and scavenging properties of (E)- and (Z)-resveratrol[J].Life Sciences,2000,66(9):769-777.
[21] PARK C K,LEE H T,CHEONG H T,et al.Effect of superoxide dismutase (SOD) on pronucleus formation of porcine oocytes fertilized in vitro[J].Theriogeno-logy,1997,48(7):1137-1146.
[22] EVA T,ANTON K,PETER M,et al.Resveratrol offers protection to oxidative stress induced by ferrous ascorbate in bovine spermatozoa[J].Journal of Environmental Science and Health,2015,50(14):1071153.
[23] 李崇阳.白藜芦醇和褪黑素对奶牛性控冻精精子质量和体外受精能力的影响[D].北京:中国农业科学院,2018. LI C Y.Effects of resveratrol and melatonin on the quality and in vitro fertilization ability of sex-sorted Holstein bull spermatozoa[D].Beijing:Chinese Academy of Agricultural Sciences,2018.(in Chinese)