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The Antioxidant Icariin Protects Porcine Oocytes against Damage during Aging In Vitro

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Alternative Title
황산화제 이카린이 체외에서 노화동안 돼지 난자의 손상으로부터 보호에 미치는 영향
Abstract
Icariin (ICA) is found in all species of Epimedium herbs, has strong antioxidant activity, and is thought to exert anti-aging effects in vitro. We investigated whether ICA treatment protects oocytes against aging in vitro. Treatment with 5 µM ICA (ICA-5) significantly decreased reactive oxygen species activity and increased the level of glutathione and mRNA expression of antioxidant genes (SOD1, SOD2, PRDX5, and NFE2L2) during aging. In addition, ICA-5 prevented defects in spindle formation and chromosomal alignment, and increased mRNA expression of cytoplasmic maturation factor genes (BMP15, CCNB1, MOS, and GDF9). ICA-5 prevented apoptosis, increased mRNA expression of anti-apoptotic genes (BCL2L1 and BIRC5), and reduced mRNA expression of pro-apoptotic genes (BAK1 and CASP3). 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. These results indicate that ICA protects porcine oocytes against damage during aging in vitro by preventing oxidative stress.
이카린 (ICA)은 모든 종의 에피메디움 허브에서 존재하며 강력한 항산화 활성을 가지며 시험관 내에서 노화 방지 효과를 발휘하는 것으로 생각된다. 우리는 ICA 처리가 시험관 내 노화에 대해 난모세포를 보호하는지 여부를 조사 하였다. 5 μM ICA (ICA-5)로 처리하면 노화 동안 활성산소 수준은 크게 감소하고 글루타티온 수준과 항산화 유전자 (SOD1, SOD2, PRDX5 및 NFE2L2)의 mRNA 발현이 증가하였다. 또한, ICA-5 는 스핀들 형성 및 염색체 정렬에서의 결함을 방지하고 세포질 성숙 인자 유전자 (BMP15, CCNB1, MOS 및 GDF9)의 mRNA 발현을 증가시켰다. ICA-5 는 세포사멸을 방지하고, 항-세포사멸 유전자 (BCL2L1 및 BIRC5)의 mRNA 발현을 증가시키고, 세포사멸 유전자 (BAK1 및 CASP3)의 mRNA 발현을 감소시켰다. 성숙 및 난할율은 모든 그룹에서 유사하지만, 배반포 당 총 세포 수 및 배반포 단계에서의 세포사멸 세포의 백분율은 각각 노화 그룹보다 대조군 및 ICA-5 그룹에서 더 높고 더 낮았다. 이러한 결과는 ICA 가 산화스트레스를 방지함으로써 시험관내 노화 동안 돼지의 난모세포를 손상으로부터 보호한다는 것을 나타낸다.
Author(s)
Yoon, Jae Wook
Issued Date
2020
Awarded Date
2020. 2
Type
Dissertation
URI
http://dcoll.jejunu.ac.kr/common/orgView/000000009319
Alternative Author(s)
윤재욱
Affiliation
제주대학교 대학원
Department
대학원 생명공학과
Advisor
박세필
Table Of Contents
CONTENTS i
LIST OF FIGURES .ⅲ
LIST OF TABLES ⅳ
ABSTRACT. 1
1. INTRODUCTION. 2
2. MATERIALS AND METHODS 5
2.1. Chemicals and reagents. 5
2.2. Aging and IVM of porcine oocyte.. 5
2.3. Oocyte aging and ICA treatment... 5
2.4. PA and embryo culture 6
2.5. Measurement of intracellular ROS and GSH levels. 6
2.6. Immunofluorescence. 7
2.7. Terminal deoxynucleotidyl transferase dUTP nick-end labeling and Hoechst staining. 8
2.8. mRNA extraction and complementary DNA synthesis 8
2.9. Real-time RT-PCR 8
2.10. Western blot analysis 11
2.11. Statistical analysis. 11
3. RESULTS. 12
3.1. ICA enhances the in vitro embryo development of aging porcine oocytes 12
3.2. ICA reduces the level of ROS in aging porcine oocytes in vitro 14
3.3. ICA prevents aberrant spindle organization and chromosome misalignment in aging porcine oocytes in vitro 16
3.4. ICA increases expression of cytoplasmic maturation markers in aging porcine oocytes in vitro 18
3.5. ICA increases expression of estrogen receptor genes in aging porcine oocytes in vitro. 20
3.6. ICA changes expression of apoptosis-related genes in aging porcine oocytes in vitro 22
3.7. ICA improves the developmental capacity and quality of embryos derived from aging porcine oocytes in vitro. 24
4. DISCUSSION. 26
REFERENCES 32
ABSTRACT IN KOREAN. 39
ACKNOWLEDGEMENT
Degree
Master
Publisher
제주대학교 대학원
Citation
Yoon, Jae Wook. (2020). The Antioxidant Icariin Protects Porcine Oocytes against Damage during Aging In Vitro
Appears in Collections:
Faculty of Biotechnology > Molecular Biotechnology
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