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Anim Biosci > Volume 34(4); 2021 > Article
Animal Reproduction and Physiology
Animal Bioscience 2021;34(4): 546-557.
DOI: https://doi.org/10.5713/ajas.20.0046    Published online May 12, 2020.
The antioxidant icariin protects porcine oocytes from age-related damage in vitro
Jae-Wook Yoon1,2,a  , Seung-Eun Lee1,2,a  , Yun-Gwi Park1,2  , Won-Jae Kim1,2  , Hyo-Jin Park1,2  , Chan-Oh Park1,2  , So-Hee Kim1,2  , Seung-Hwan Oh1,2  , Do-Geon Lee1,2  , Da-Bin Pyeon1,2  , Eun-Young Kim1,2,3  , Se-Pill Park1,2,3,* 
1Faculty of Biotechnology, College of Applied Life Sciences, Jeju National University, Jeju 63243, Korea
2Stem Cell Research Center, Jeju National University, Jeju 63243, Korea
3Mirae Cell Bio, Seoul 04795, Korea
Correspondence:  Se-Pill Park, Tel: +82-64-754-4650, Fax: +82-0303-3130-4650, Email: sppark@jejunu.ac.kr
Received: 28 January 2020   • Revised: 26 February 2020   • Accepted: 29 April 2020
aThese authors contributed equally to this work.
Abstract
Objective
If fertilization does not occur within a specific period, the quality of unfertilized oocytes in the oviduct (in vivo aging) or in culture (in vitro aging) will deteriorate over time. Icariin (ICA), found in all species of Epimedium herbs, has strong antioxidant activity, and is thought to exert anti-aging effects in vitro. We asked whether ICA protects oocytes against age-related changes in vitro.
Methods
We analyzed the reactive oxygen species (ROS) levels and expression of antioxidant, maternal, and estrogen receptor genes, and along with spindle morphology, and the developmental competence and quality of embryos in the presence and absence of ICA.
Results
Treatment with 5 μM ICA (ICA-5) led to a significant reduction in ROS activity, but increased mRNA expression of glutathione and antioxidant genes (superoxide dismutase 1 [SOD1], SOD2, peroxiredoxin 5, and nuclear factor erythroid 2‐like 2), during aging in vitro. In addition, ICA-5 prevented defects in spindle formation and chromosomal alignment, and increased mRNA expression of cytoplasmic maturation factor genes (bone morphogenetic protein 15, cyclin B1, MOS proto‐oncogene, serine/threonine kinase, and growth differentiation factor‐9). It also prevented apoptosis, increased mRNA expression of antiapoptotic genes (BCL2-like 1 and baculoviral IAP repeat-containing 5), and reduced mRNA expression of pro-apoptotic genes (BCL2 antagonist/killer 1 and activation of caspase-3). Although the maturation and cleavage rates were similar in all groups, the total cell number per blastocyst and the percentage of apoptotic cells at the blastocyst stage were higher and lower, respectively, in the control and ICA-5 groups than in the aging group.
Conclusion
ICA protects oocytes against damage during aging in vitro; therefore, it can be used to improve assisted reproductive technologies.
Keywords: In vitro Aging; Porcine; Oocyte; Icariin; Antioxidant


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