• 한국수정란이식학회지
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• 제31권3호
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• pp.153-159
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• 2016

(-)-Epicatechin gallate (ECG) is a polyphenol compound of green tea exhibiting biological activities, such as antioxidant and anticancer effects. To examine the effect of ECG on porcine oocytes during in vitro maturation (IVM), oocytes were treated with 0-, 5-, 15-, and $25{\mu}M$ ECG. After maturation, we investigated nuclear maturation, intracellular glutathione (GSH) and reactive oxygen species (ROS) levels and subsequent embryonic development after parthenogenetic activation (PA) and in vitro fertilization (IVF). After 42 hours of IVM, the $5{\mu}M$ group exhibited significantly increased (p< 0.05) nuclear maturation (89.8%) compared with the control group (86.1%). However, the $25{\mu}M$ group observed significantly decreased (p< 0.05) nuclear maturation (83.5%). In intracellular maturation assessment the 5-, 15-, and $25{\mu}M$ groups had significantly increased (p< 0.05) GSH levels and decreased ROS levels compared with the controls. The 5- and $15{\mu}M$ group showed significantly increased (p< 0.05) embryo formation rates and total cell number of blastocysts after PA (18% and 68.9, 15% and 85.1 vs. 12% and 59.5, respectively) compared with controls. Although the $25{\mu}M$ group observed significantly lower blastocyst formation rates after PA (27.6% vs. 23.2%) than control group, the $5{\mu}M$ group showed significantly increased blastocyst formation rates after PA (37.2% vs. 23.2%) compared to the control group. Furthermore, the $5{\mu}M$ group measured significantly increased blastocyst formation rates (20.7% vs. 8.6%) and total cell number after IVF ($88.3{\pm}1.5$ vs. $58.0{\pm}3.6$) compared to the control group. The treatment of $5{\mu}M$ ECG during IVM affectively improved the porcine embryonic developmental competence by regulating intracellular oxidative stress during IVM.

• Asian-Australasian Journal of Animal Sciences
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• 제29권8호
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• pp.1095-1101
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• 2016

Ginsenoside Rg1 is a natural compound with various efficacies and functions. It has beneficial effects on aging, diabetes, and immunity, as well as antioxidant and proliferative functions. However, its effect on porcine embryo development remains unknown. We investigated the effect of ginsenoside Rg1 on the in vitro development of preimplantation porcine embryos after parthenogenetic activation in high-oxygen conditions. Ginsenoside treatment did not affect cleavage or blastocyst formation rates, but did increase the total cell number and reduced the rate of apoptosis. In addition, it had no effect on the expression of four apoptosis-related genes (Bcl-2 homologous antagonist/killer, B-cell lymphoma-extra large, Caspase 3, and tumor protein p53) or two metabolism-related genes (mechanistic target of rapamycin, carnitine palmitoyltransferase 1B), but increased the expression of Glucose transporter 1 (GLUT1), indicating that it may increase glucose uptake. In summary, treatment with the appropriate concentration of ginsenoside Rg1 ($20{\mu}g/mL$) can increase glucose uptake, thereby improving the quality of embryos grown in high-oxygen conditions.

• 한국응용과학기술학회지
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• 제36권1호
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• pp.258-268
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• 2019

The extracts of AS contain in alloxanthin, halocynthiaxanthin, astaxanthin and 13 kinds of carotenoids. The aim of the study was to assess the anti-oxidant activity and cell viability of AS. The anti-oxidant activity was determined by using DPPH radical inhibition activity and superoxide dismutase (SOD)-like activity. The results of cell viability assay showed that the extracts from AS were cytotoxic at concentrations above $5.0mg/m{\ell}$. This study was designed to examine inflammation induced by LPS, protection effect by UVB and the toxicity of Ascidian shell extract(ASE) as a functional cosmetic ingredient. Evaluation of embryo toxicity resulted in embryo coagulation and mortality when treated at 5.0, 10.0, $20.0mg/m{\ell}$. At the lowest concentration of $1.0mg/m{\ell}$, hatchability resulted in 100.0 % rate. The results of arrhythmia measurement in larvae showed similarity to the evaluation of embryo toxicity. This result demonstrated that toxicity is present at concentrations greater than $5.0mg/m{\ell}$. The protective effect of ASE on LPS and UVB-induced in the zebrafish was investigated. Intracellular reactive oxygen species(ROS) generated by the exposure of zebrafish to LPS, UVB-radiation were significantly decreased after treatment with ASE at $0.1mg/m{\ell}$. As a result, ASE similarly reduced UVB-induced ROS generation and cell death in live zebrafsih. Therefore, it is suggested that ASE has anti-Inflammatory effects and can possibly be used as a functional substance for skin protection in the future.

• 한국수정란이식학회지
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• 제32권4호
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• pp.257-265
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• 2017

Ferulic Acid (FA) is a metabolite of phenylalanine and tyrosine, a phenolic compound commonly found in fruits and vegetables. Several studies have shown that FA has various functions such as antioxidant effect, prevention of cell damage from irradiation, protection from cell damage caused by oxygen deficiency, anti-inflammatory action, anti-aging action, liver protective effect and anti-cancer action. In this study, we investigated the maturation rate, intracellular glutathione (GSH) and reactive oxygen species (ROS) of porcine oocytes by adding FA to the in vitro maturation (IVM) medium and examined subsequent embryonic developmental competence at 5% oxygen through parthenogenesis. There is no significant difference between the control group ($0{\mu}M$) and treatment groups ($5{\mu}M$, $10{\mu}M$, $20{\mu}M$) on maturation rates. Intracellular GSH levels in oocyte treated with $5{\mu}M$ of FA significantly increased (P < 0.05), and $20{\mu}M$ of FA revealed significant decrease (P < 0.05) in intracellular ROS levels compared with the control group. Oocytes treated with FA exhibited significantly higher cleavage rates (79.01% vs 89.19%, 92.20%, 90.89%, respectively) than the control group. Oocytes treated with $10{\mu}M$ showed significantly higher blastocyst formation rates (28.3% vs 40.3%, respectively) after PA than the control group. Total cell numbers in blastocyst of $10{\mu}M$ FA displayed significantly higher (39.4 vs 51.9, respectively) than the control group. In conclusion, these results suggested that treatment with FA during IVM improved the developmental potential of porcine embryos by increasing intracellular GSH synthesis and reducing ROS levels. Also, there was an improvement of cleavage rate, blastocyst formation and total cell numbers in blastocysts. It might be associated with Keap1-Nrf2 pathway as an antioxidant regulate pathway that plays a crucial role in determining the sensitivity of cells to oxidative damages by regulating the basal and inducible expression of enzymes which is related to detoxification and anti-oxidative effects, stress response enzymes and/or proteins and ABC transporters.

• 한국수정란이식학회지
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• 제30권3호
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• pp.121-128
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• 2015

The objective of this study was to evaluate the efficiency of sperm cryosurvival in boar sperm separated by Percoll containing antioxidant enzymes. The boar semen was collected into a pre-warmed ($37^{\circ}C$) thermos bottle by gloved-hand method and was separated by 65% Percoll with superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) before freezing. The frozen sperm was thawed at $38.5^{\circ}C$ for 45 sec in water-bath for sperm characteristic analysis. The sperm were estimated with SYBR14/PI double staining for viability, FITC-PNA/PI double staining for acrosome reaction, Rhodamine123/PI double staining for mitochondrial integrity and were analyzed using flow cytometry. In results, sperm viability, acrosome reaction and mitochondrial integrity were improved in separated sperm groups compared with unseparated sperm by Percoll (UP) group. Especially, viability was significantly higher in sperm separated by Percoll containing 400 IU CAT group compared with other groups (P<0.05). And acrosome reaction was decreased in sperm separated by Percoll with 300 IU SOD, 400 IU CAT and 0.5 mM GSH groups compared with other groups, however, there were no significantly difference mitochondrial integrity among sperm separated by Percoll with antioxidant enzymes. In conclusion, we suggest that use of Percoll containing antioxidant enzymes for sperm separation will be beneficial for sperm cryopreservation in pigs.

• Clinical and Experimental Reproductive Medicine
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• 제49권2호
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• pp.117-126
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• 2022

Objective: This study examined whether the addition of triple antioxidants (3A)-10 µM acetyl-L-carnitine, 10 µM N-acetyl-L-cysteine, and 5 µM α-lipoic acid-in freezing-thawing medium during human sperm cryopreservation using the sucrose vitrification (SuV) and liquid nitrogen vapor (Vapor) techniques could improve post-thaw survival of spermatozoa. Methods: We analyzed 30 samples from healthy human sperm donors. Each sample was allocated into one of five groups: fresh control, SuV, SuV+3A, Vapor, and Vapor+3A. The sperm motility, morphology, viability, intracellular and extracellular reactive oxygen species (ROS) levels, and sperm DNA fragmentation (SDF) were evaluated. Results: The cryopreserved spermatozoa had significantly reduced percentages of motility (p<0.05) and viability (p<0.05). Antioxidant supplementation non-significantly improved these parameters (p>0.05). No significant differences were found in sperm morphology between the fresh and frozen-thawed groups (p>0.05). After freezing, the extracellular ROS levels in the frozen-thawed groups were significantly higher (p<0.05) than in the fresh group. However, we did not find any differences in intracellular ROS parameters among these groups (p>0.05). The SDF was higher in the SuV and Vapor groups than in the fresh group, but without statistical significance (p=0.075 and p=0.077, respectively). Conclusion: Cryopreservation had detrimental effects on sperm motility, viability, and extracellular ROS levels, without changing the morphology or intracellular ROS levels. Antioxidant supplementation was slightly effective in preventing SDF in frozen-thawed spermatozoa.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2002년도 춘계학술발표대회 발표논문초록집
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• pp.46-46
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• 2002

본 연구는 돼지의 미성숙 난포란을 체외에서 성숙, 수정시킨 후, 생산된 체외수정란을 NCSU 23 체외배양액에 항산화제 (NAC, ebselen 및 glutathione) 와 glutathione 합성억제제인 BSO의 첨가가 돼지 체외 수정란의 체외 발육에 미치는 영향을 검토하였다. Glutathione 합성 억제제인 BSO가 돼지 수정란의 체외 발육에 미치는 영향을 검토한 결과, 0, 1.0 및 5.0mM의 BSO 을 첨가하여 상실배기 이상 발육된 체외 발육 성적은 각각 35.9, 15.7 및 8.4%로서 BSO 첨가구가 대조구에 비해 통계적으로 유의하게 낮은 체외배율성적을 나타냈으며 (P＜0.05), NAC 1.0mM, ebselen 10μM 및 glutathione 100μM 과 BSO 1.0 mM 을 각각 첨가하여 체외 배양을 실시한 경우, 상실배 이상 발육된 체외 발육율은 40.5, 44.2, 36.0 및 10.9%로서 항산화제 첨가구가 BSO 첨가구보다 통계적으로 유의하게 높은 체외 발육 성적을 나타냈다 (P＜0.05). 모든 처리구에서 체외 수정 후 생산된 배반포기 수정란의 세포수에는 차이가 인정되지 않았다. (중략)

• 한국응용과학기술학회지
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• 제38권1호
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• pp.118-125
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• 2021

본 연구에서 사용된 와송(Orostachys japonica A. Berger)은 암 치료를 위한 민간요법으로 오랫동안 사용되어 온 약초이다. 본 연구에서는 와송(Orostachys japonica A. Berger) 에틸아세테이트 분획물의 항산화 효능 활성에 대하여 실험한 결과를 보고한다. 와송(Orostachys japonica A. Berger) 에틸아세테이트 분획물의 항산화 효능은 0.10 mg/mL 농도에서 78.54%의 DPPH 라디칼 소거활성을 보였고, 73.48%의 ABTS+ 라디칼 소거활성을 나타내어 항산화 효과가 있는 것을 확인하였다. 또한 대체실험동물 모델인 제브라피쉬를 이용한 독성실험 결과에서는 모든 농도에서 100%의 배아 생존율을 보이고 응고 또는 부화지연을 관찰할 수 없어 독성이 없는 것으로 나타났으며, UV-B조사에 의해 유도된 ROS생성은 와송 에틸아세테이트 분획물을 처리한 모든 larvae에서 전체적으로 형광강도가 감소하였다. 특히 3 ㎍/mL 농도에서 양성대조군 대비 35.7%의 감소율을 보여 효과적으로 ROS 생성을 억제하였음을 확인하였다. 위와 같은 결과로 미루어보아, 와송 에틸아세테이트 분획물은 천연 항산화제로서 화장품 분야에서 응용 가능성이 있을 것으로 기대한다.

• 한국수정란이식학회지
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• 제26권4호
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• pp.257-263
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• 2011

BPA, a diphenyl compound containing groups, that make it structurally similar to synthetic estrogen and is considered as one of the major endocrine disruptors. Silymarin has extensively been used to prevent and/or alleviate some human disease, especially for the treatment of adverse liver conditions. It has an antioxidative efficacy and cancer preventive efficacy. Therefore, we examined the hypothesis that silymarin can inhibit BPA-induced toxicity in boar sperm duing in vitro storage. Sperm characteristics (motility, viability, membrane integrity and mitochondrion activity) in semen exposed to BPA (10~200 uM) were sharply lowered, while it increase in a dose and time dependent manner due to silymarin addition (50~200 uM) into semen extender in the presence of BPA (100 uM). All of the evaluated characteristics were gradually improved in the groups that were treated with silymarin (50~200 uM) in the presence of BPA (100 uM) in comparison to BPA 100 uM alone group, irrespective of incubation periods (3 and 6 h). These results demonstrate that silymarin can ameliorate the toxicity of BPA on boar sperm characteristics during in vitro storage, suggesting that silymarin indirectly act as an antioxidant.

• 한국수정란이식학회지
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• 제19권1호
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• pp.67-73
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• 2004

본 연구는 우리나라 고유의 유전자원인 재래산양의 체내수정란 생산기술을 확립하고자 수행하였다. 흡입법(aspiration)과 세절법(slicing)에 의해 난소 한 개당 회수된 난자의 수는 3.9개와 4.1개를 나타내어 slicing방법이 aspiration방법보다는 많은 숫자의 난자를 회수하였으나 유의적인 차이는 나타내지 않았다. 회수된 난자의 등급별 분포는 aspiration방법에서 Grade I, Grade II, Grade III, Grade IV의 비율이 10.3%, 20.5%, 38.5%, 30.8%를 나타내었으며, slicing법에서는 9.8%, 22.0%, 39.0%, 29.3%를 나타내어 Grade III과 Grade IV의 비율이 70% 이상을 차지하였다. 회수된 난자를 체외성숙 시킨 결과 Grade I과 Grade II에서는 85% 이상이 metaphase H (MII)까지 도달하였으나, Grade III과 Grade IV는 40% 이하의 체외성숙율을 나타내었다. 체외수정용 배양액으로 BO를 사용하였을 경우 Grade I 및 II에서 84.4%의 난분할율을 나타내어 TALP를 사용하였을 때의 58.8%보다 높은 난할율을 보였다. 또한 배양액의 종류별 체외발달율에 있어서는 상실배 및 배반포기배로의 발달은 mSOF를 배양액으로 이용하였을 경우 15.0%의 발달율을 나타내었으며, 체외배양 시 항산화물질인 glutathione (GSH)을 첨가함으로서 26.8%의 상실배 및 배반포배로의 발달율을 나타내었다.