Effects of epigallocatechin-3-gallate on bovine oocytes matured in vitro

  • Huang, Ziqiang (Embryo Biotechnology and Reproduction Laboratory, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences) ;
  • Pang, Yunwei (Embryo Biotechnology and Reproduction Laboratory, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences) ;
  • Hao, Haisheng (Embryo Biotechnology and Reproduction Laboratory, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences) ;
  • Du, Weihua (Embryo Biotechnology and Reproduction Laboratory, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences) ;
  • Zhao, Xueming (Embryo Biotechnology and Reproduction Laboratory, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences) ;
  • Zhu, Huabin (Embryo Biotechnology and Reproduction Laboratory, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences)
  • Received : 2017.12.07
  • Accepted : 2018.03.05
  • Published : 2018.09.01


Objective: Epigallocatechin-3-gallate (EGCG) is a major ingredient of catechin polyphenols and is considered one of the most promising bioactive compounds in green tea because of its strong antioxidant properties. However, the protective role of EGCG in bovine oocyte in vitro maturation (IVM) has not been investigated. Therefore, we aimed to study the effects of EGCG on IVM of bovine oocytes. Methods: Bovine oocytes were treated with different concentrations of EGCG (0, 25, 50, 100, and $200{\mu}M$), and the nuclear and cytoplasmic maturation, cumulus cell expansion, intracellular reactive oxygen species (ROS) levels, total antioxidant capacity, the early apoptosis and the developmental competence of in vitro fertilized embryos were measured. The mRNA abundances of antioxidant genes (nuclear factor erythriod-2 related factor 2 [NRF2], superoxide dismutase 1 [SOD1], catalase [CAT], and glutathione peroxidase 4 [GPX4]) in matured bovine oocytes were also quantified. Results: Nuclear maturation which is characterized by first polar body extrusion, and cytoplasmic maturation characterized by peripheral and cortical distribution of cortical granules and homogeneous mitochondrial distribution were significantly improved in the $50{\mu}M$ EGCG-treated group compared with the control group. Adding $50{\mu}M$ EGCG to the maturation medium significantly increased the cumulus cell expansion index and upregulated the mRNA levels of cumulus cell expansion-related genes (hyaluronan synthase 2, tumor necrosis factor alpha induced protein 6, pentraxin 3, and prostaglandin 2). Both the intracellular ROS level and the early apoptotic rate of matured oocytes were significantly decreased in the $50{\mu}M$ EGCG group, and the total antioxidant ability was markedly enhanced. Additionally, both the cleavage and blastocyst rates were significantly higher in the $50{\mu}M$ EGCG-treated oocytes after in vitro fertilization than in the control oocytes. The mRNA abundance of NRF2, SOD1, CAT, and GPX4 were significantly increased in the $50{\mu}M$ EGCG-treated oocytes. Conclusion: In conclusion, $50{\mu}M$ EGCG can improve the bovine oocyte maturation, and the protective role of EGCG may be correlated with its antioxidative property.


Epigallocatechin-3-gallate;Reactive Oxygen Species (ROS);Oocytes;Oxidative Stress;Antioxidant;Bovine


  1. Flores D, Souza V, Betancourt M, et al. Oxidative stress as a damage mechanism in porcine cumulus-oocyte complexes exposed to malathion during in vitro maturation. Environ Toxicol 2017;32:1669-78.
  2. Tsunoda S, Kimura N, Fujii J. Oxidative stress and redox regulation of gametogenesis, fertilization, and embryonic development. Reprod Med Biol 2014;13:71-9.
  3. de Lamirande E, Lamothe G. Reactive oxygen-induced reactive oxygen formation during human sperm capacitation. Free Radic Biol Med 2009;46:502-10.
  4. Agarwal A, Saleh RA, Bedaiwy MA. Role of reactive oxygen species in the pathophysiology of human reproduction. Fertil Steril 2003;79:829-43.
  5. Luderer U. Ovarian toxicity from reactive oxygen species. Vitam Horm 2014;94:99-127.
  6. Gonzalez-Fernandez R, Hernandez J, Martin-Vasallo P, et al. Expression levels of the oxidative stress response gene ALDH3A2 in granulosa-lutein cells are related to female age and infertility diagnosis. Reprod Sci 2016;23:604-9.
  7. Roychoudhury S, Agarwal A, Virk G, Cho CL. Potential role of green tea catechins in the management of oxidative stressassociated infertility. Reprod Biomed Online 2017;34:487-98.
  8. Hijazi MM, Khatoon N, Azmi MA, et al. Report: effects of Camellia sinensis L. (green tea) extract on the body and testicular weight changes in adult Wistar rate. Pak J Pharm Sci 2015;28:249-53.
  9. Rahmani AH, Al Shabrmi FM, Allemailem KS, Aly SM, Khan MA. Implications of green tea and its constituents in the prevention of cancer via the modulation of cell signalling pathway. Biomed Res Int 2015;2015:Article ID 925640.
  10. Xu Q, Langley M, Kanthasamy AG, Reddy MB. Epigallocatechin gallate has a neurorescue effect in a mouse model of parkinson disease. J Nutr 2017;147:1926-31.
  11. Spinaci M, Volpe S, De Ambrogi M, Tamanini C, Galeati G. Effects of epigallocatechin-3-gallate (EGCG) on in vitro maturation and fertilization of porcine oocytes. Theriogenology 2008; 69:877-85.
  12. Vanderhyden BC, Caron PJ, Buccione R, Eppig JJ. Developmental pattern of the secretion of cumulus expansion-enabling factor by mouse oocytes and the role of oocytes in promoting granulosa cell differentiation. Dev Biol 1990;140:307-17.
  13. Pang YW, Sun YQ, Sun WJ, et al. Melatonin inhibits paraquatinduced cell death in bovine preimplantation embryos. J Pineal Res 2016;60:155-66.
  14. Kothari S, Thompson A, Agarwal A, du Plessis SS. Free radicals: Their beneficial and detrimental effects on sperm function. Indian J Exp Biol 2010;48:425-35.
  15. Ufer C, Wang CC, Borchert A, Heydeck D, Kuhn H. Redox control in mammalian embryo development. Antioxid Redox Signal 2010;13:833-75.
  16. Tiwari M, Chaube SK. Moderate increase of reactive oxygen species triggers meiotic resumption in rat follicular oocytes. J Obstet Gynaecol Res 2016;42:536-46.
  17. Wang ZG, Yu SD, Xu ZR. Improvement in bovine embryo production in vitro by treatment with green tea polyphenols during in vitro maturation of oocytes. Anim Reprod Sci 2007; 100:22-31.
  18. Roth Z, Aroyo A, Yavin S, Arav A. The antioxidant epigallocatechin gallate (EGCG) moderates the deleterious effects of maternal hyperthermia on follicle-enclosed oocytes in mice. Theriogenology 2008;70:887-97.
  19. Li Y, Zhang C, Gao Y, et al. Effect of epigallocatechin-3-gallate on the in vitro developmental potential of porcine oocytes and embryos obtained parthenogenetically and by somatic cell nuclear transfer. Italian J Anim Sci 2016;13:3116.
  20. Pisoschi AM, Pop A. The role of antioxidants in the chemistry of oxidative stress: a review. Eur J Med Chem 2015;97:55-74.
  21. Meng Q, Velalar CN, Ruan R. Effects of epigallocatechin-3-gallate on mitochondrial integrity and antioxidative enzyme activity in the aging process of human fibroblast. Free Radic Biol Med 2008;44:1032-41.
  22. Sahin K, Orhan C, Tuzcu M, et al. Epigallocatechin-3-gallate prevents lipid peroxidation and enhances antioxidant defense system via modulating hepatic nuclear transcription factors in heat-stressed quails. Poult Sci 2010;89:2251-8.
  23. Han XD, Zhang YY, Wang KL, et al. The involvement of Nrf2 in the protective effects of (-)-Epigallocatechin-3-gallate (EGCG) on NaAsO2-induced hepatotoxicity. Oncotarget 2017;8:65302-12.
  24. Dinkova-Kostova AT, Abramov AY. The emerging role of Nrf2 in mitochondrial function. Free Radic Biol Med 2015;88:179-88.
  25. Sriram N, Kalayarasan S, Sudhandiran G. Epigallocatechin-3-gallate augments antioxidant activities and inhibits inflammation during bleomycin-induced experimental pulmonary fibrosis through Nrf2-Keap1 signaling. Pulm Pharmacol Ther 2009;22:221-36.