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Protodioscin protects porcine oocytes against H2O2-induced oxidative stress during in vitro maturation |
So-Hee Kim1,2,a 
, Seung-Eun Lee1,2,a
, Jae-Wook Yoon1
, Hyo-Jin Park1
, Seung-Hwan Oh1
, Do-Geon Lee1
, Da-Bin Pyeon1
, Eun-Young Kim1,2,3
, Se-Pill Park1,3,4,*
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1Stem Cell Research Center, Jeju National University, Jeju 63243, Korea
2Faculty of Biotechnology, College of Applied Life Sciences, Jeju National University, Jeju 63243, Korea
3Mirae Cell Bio, Seoul, 04795, Korea
4Department of Bio Medical Informatic, College of Applied Life Sciences, Jeju National University, Jeju 63243, Korea |
Correspondence:
Se-Pill Park, Tel: +82-64-754-4650, Fax: +82-0303-3130-4650, Email: sppark@jejunu.ac.kr
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Received: 20 July 2022 • Revised: 19 September 2022 • Accepted: 7 November 2022
aThese authors contributed equally to this work. |
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| Abstract |
Objective
The present study investigated whether protodioscin (PD), a steroidal saponin mainly found in rhizome of Dioscorea species, alleviates oxidative stress-induced damage of porcine oocytes during in vitro maturation.
Methods
Oocytes were treated with different concentrations of PD (0, 1, 10, 100, and 200 μM) in the presence of 200 μM H2O2 during in vitro maturation. Following maturation, spindle morphology and mitogen-activated protein kinase activity was assessed along with reactive oxygen species level, GSH activity, and mRNA expression of endogenous antioxidant genes at the MII stage. On the day 7 after parthenogenetic activation, blastocyst formation rate was calculated and the quality of embryo and mRNA expression of development-related genes was evaluated.
Results
Developmental competence was significantly poorer in the 0 μM PD-treated (control) group than in the non-treated (normal) and 10 μM PD-treated (10PD) groups. Although the reactive oxygen species level did not significantly differ between these three groups, the glutathione level and mRNA expression of antioxidant genes (superoxide dismutase 1 [SOD1], SOD2, nuclear factor erythroid 2-related factor 2 [Nrf2], and hemo oxygenase-1 [HO-1]) were significantly higher in the normal and 10PD groups than in the control group. In addition, the percentage of oocytes with defective spindle and abnormal chromosomal alignment was significantly lower and the ratio of phosphorylated p44/42 to total p44/42 was significantly higher in the normal and 10PD groups than in the control group. The total cell number per blastocyst was significantly higher in the 10PD group than in the control group. The percentage of apoptotic cells in blastocysts was highest in the control group; however, the difference was not significant. mRNA expression of development-related genes (POU domain, class 5, transcription factor 1 [POU5F1], caudal type homeobox 2 [CDX2], Nanog homeobox [NANOG]) was consistently increased by addition of PD.
Conclusion
The PD effectively improves the developmental competence and quality of blastocysts by protecting porcine oocytes against oxidative stress. |
| Keywords:
Antioxidant; In vitro Maturation; Oxidative Stress; Protodioscin; Reactive Oxygen Species (ROS) |
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