[논문]Effects of Antioxidants Supplement in Porcine Sperm Freezing on in vitro Fertilization and the Glutathione and Reactive Oxygen Species Level of Presumptive Zygotes

Park, Sang-Hyoun; Jeon, Yubyoel; Yu, Il-Jeoung

doi:10.12750/jet.2017.32.4.337

Effects of Antioxidants Supplement in Porcine Sperm Freezing on in vitro Fertilization and the Glutathione and Reactive Oxygen Species Level of Presumptive Zygotes 원문보기

Journal of embryo transfer = 한국수정란이식학회지, v.32 no.4, 2017년, pp.337 - 342

Park, Sang-Hyoun (Department of Theriogenology and Reproductive Biotechnology, College of Veterinary Medicine and Bio-safety Research Institute, Chonbuk National University) , Jeon, Yubyoel (Department of Theriogenology and Reproductive Biotechnology, College of Veterinary Medicine and Bio-safety Research Institute, Chonbuk National University) , Yu, Il-Jeoung (Department of Theriogenology and Reproductive Biotechnology, College of Veterinary Medicine and Bio-safety Research Institute, Chonbuk National University)

Abstract ▼ AI-Helper

The present study was aimed to determine the effects of green tea extract (GTE) and beta-mercaptoethanol (${\beta}-ME$) supplementation in boar sperm freezing extender on in vitro fertilization (IVF) and reactive oxygen species (ROS) and glutathione (GSH) levels of presumptive zygotes (PZs). Experimental groups were allocated into lactose egg yolk (LEY) without antioxidant (control), GTE (1,000 mg/l in LEY) and ${\beta}-ME$ ($50{\mu}M$ in LEY). In freezing, spermatozoa extended with LEY were cooled to $5^{\circ}C$ for 3 h and then kept at $5^{\circ}C$ for 30 min following dilution with LEY containing 9% glycerol and 1.5% Equex STM. The final sperm concentration was $1{\times}10^8/ml$. Spermatozoa were loaded into straws and frozen in nitrogen vapor for 20 min. For IVF, oocytes were matured in NCSU-23 medium and co-cultured with spermatozoa following thawing at $37^{\circ}C$ for 25 sec. At 12 h following IVF, IVF parameters (sperm penetration and monospermy) were evaluated. In addition, GSH and ROS levels of PZs were determined by Cell Tracker Blue CMF2HC and DCHFDA, respectively. IVF parameters did not show any significant difference among the experimental groups. GSH and ROS levels of PZs were not significantly different between groups. In conclusion, antioxidant supplementation in boar sperm freezing could not influence IVF parameters, ROS and GSH levels of PZs.

주제어

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제안 방법

The objectives of the present study were aimed to determine the effects of GTE and β-ME supplementation in boar sperm freezing extender on IVF parameters and ROS and GSH levels of presumptive zygotes (PZs).
Therefore, we will evaluate the effect of addition of GTE and β-ME in post-thawed spermatozoa to know the latent antioxidant effect on IVF parameters and ROS and GSH level of presumptive zygote.

대상 데이터

Eight boars (2 Duroc jersey and 6 Yorkshire) were used for this experiment. The semen collected using gloved-hand technique was filtered through four layers of sterile gauze to remove the gel particle.
Five replicates were conducted for each experiment. Percentage data were subjected to arcsin transformation before analysis.
Green tea powder (GTP) was obtained from Bioland (GTP-020968, Seoul, Korea). GTE and the β-ME stock were prepared by modifying the stock prepared by Park and Yu (2017).

데이터처리

All data are presented as means ± SE and were analyzed using ANOVA followed by Duncan’s multiple range test.

성능/효과

It is concluded that there is no positive effect of supplementation of GTE and β-ME in freezing extender on IVF parameters and ROS and GSH levels of PZs.
Likewise, as the results of IVF parameters, GSH and ROS levels of PZs following IVF with boar spermatozoa cryopreserved with GTE or β-ME or without GTE and β-ME had no significant difference among the groups.
Briefly, the presumptive zygotes were mounted on slides, fixed for 7 days with acetic acid in ethanol (1:3, v:v) at room temperature, stained with aceto-orcein [1% (w:v) orcein in 45% (v:v) acetic acid] and destained with glycerol:acetic acid:DW (1:1:3, v:v) and examined under a light microscope at ×400 magnification. The fertilization parameters evaluated were: penetration (percentage of the number of penetrated oocytes/total inseminated) and monospermy (percentage of the number of monospermic oocytes/total penetrated).

참고문헌 (28)

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원문보기 상세보기
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원문보기 상세보기
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연합인증

Effects of Antioxidants Supplement in Porcine Sperm Freezing on in vitro Fertilization and the Glutathione and Reactive Oxygen Species Level of Presumptive Zygotes 원문보기

Abstract ▼ AI-Helper

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연합인증

Effects of Antioxidants Supplement in Porcine Sperm Freezing on in vitro Fertilization and the Glutathione and Reactive Oxygen Species Level of Presumptive Zygotes 원문보기

Abstract ▼ AI-Helper

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제안 방법

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참고문헌 (28)

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