• Reproductive and Developmental Biology
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• 제40권2호
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• pp.15-22
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• 2016

As a one of unsaturated fatty acid, polyunsaturated fatty acids (PUFAs) have multiple actions: as precursor of prostaglandins (PGs), steroid hormone synthesis and energy production in animal reproduction. PUFAs, which include omega-3 (n-3) and omega-6 (n-6), are derived from the diet and changed by diet, species, breed and season. The plasma membrane of spermatozoa in mammals contain various PUFAs. These composition of PUFAs regulate the membrane fluidity and cause lipid peroxidation via generation of reactive oxygen species (ROS). Induced lipid peroxidation by ROS decreased viability and motility of spermatozoa, and it is reduced by addition of antioxidant and low concentration of PUFAs. Because oocytes of animal have a high lipid components, process of oocyte maturation and embryo development are influenced by PUFAs. In in vitro study, oocyte maturation, embryo development, intracellular cAMP and MAPK activity were increased by treatment of n-3 ${\alpha}$-linolenic acid (ALA) during maturation, whereas n-6 linoleic acid (LA) negatively influenced. Also, inhibition of fatty acid metabolism in oocyte influenced blastocyst formation of cattle. PGs are synthesized from PUFAs and various PUFAs influence PGs via regulation of PG-endoperoxide synthase (PTGS). Steroid hormone synthesis from cholesterol is regulated by expression of steroid acute regulator (StAR) protein and mRNA. Exogenous n-3 and n-6 PUFAs altered sex hormone in animal through stimulate or inhibit StAR activity. Because PUFAs altered PG and steroid hormone synthesis, follicular development was influenced by PUFAs. This effect of unsaturated fatty acid could provide information for improvement of reproductive ability in animals.

• 한국수정란이식학회지
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• 제30권1호
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• pp.23-31
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• 2015

Catalpol, an iridoid glucoside, isolated from the root of Rehmannia glutinosa Libosch. It possesses a broad range of biological and pharmacological activity including anti-tumor, anti-inflammation and anti-oxidant by acting as a free radical scavenger. Therefore, in this study, the effects of catalpol on blastocyst development, expression levels of reactive oxygen species (ROS) and apoptotic index were investigated in porcine embryos. After in vitro maturation and fertilization, porcine embryos were cultured for 6 days in porcine zygote medium 3 (PZM-3) supplemented with catalpol (0, 100, 200 and $400{\mu}M$, respectively). Blastocyst development not significantly improved in the catalpol treated group when compared with control group. Otherwise, the intracelluar levels of ROS were decreased and the numbers of apoptotic nuclei were reduced in the catalpol ($100{\mu}M$) treated porcine blastocysts (P<0.05). On the other hand, blastocyst development was significantly improved in the catalpol ($100{\mu}M$) treated group when compared with the untreated catalpol group under $H_2O_2$ ($200{\mu}M$) induced oxidative stress (P<0.05). Otherwise, the intracellular levels of ROS in catalpol ($100{\mu}M$) treated group were significantly decreased in the untreated catalpol group under $H_2O_2$ ($200{\mu}M$) induced oxidative stress (P<0.05). Furthermore, the total cell numbers of blastocysts were significantly increased (P<0.05) in the catalpol ($100{\mu}M$) treated group under $H_2O_2$ ($200{\mu}M$) induced oxidative stress, whereas numbers of apoptoic nuclei were significantly reduced (P<0.05). In conclusion, our results indicate that treatment of catalpol may have important implications for improving developmental competence and preimplantation quality of porcine embryos through its anti-oxidant and anti-apoptotic effect.

• 한국수정란이식학회지
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• 제32권3호
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• pp.249-255
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• 2017

In this experiment, we determined the effect of curcumin supplementation in freezing buffer for miniature pig sperm cryopreservation. Each ejaculate was diluted with modified Modena B extender and mixed with lactose-egg yolk (LEY extender, 80% v/v lactose solution [310 mM], 20% v/v egg yolk, and $100{\mu}g/mL$ kanamycin sulfate) and LEY-glycerol Orvus ES Paste (LEYGO, 89.5% v/v LEY, 5% v/v glycerol, 1.5% v/v Orvus ES Paste), 100 mM trehalose supplemented with 0, 10, 50, 100, and $500{\mu}M$ of curcumin from turmeric, respectively. Following equilibration, the 0.5 mL French straws were frozen and plunged into $LN_2$ tank for 7 days at least. Sperm parameter and oxidative byproducts were determined by the computer assisted sperm motility analysis (CASA) and fluorescence-activated cell sorting (FACS) as compared with each groups. Supplementation of curcumin had no effect on sperm motility, progressive motility and curvilinear velocity. However, average-path velocity and straight-line velocity were significantly higher in $10{\mu}M$ curcumin group ($100.9{\pm}8.8{\mu}m/s$, $61.7{\pm}2.9{\mu}m/s$, respectively) than control group ($77.8{\pm}3.9{\mu}m/s$, $46.4{\pm}3.0{\mu}m/s$, respectively) (p < 0.05). In addition, the level of the O2 radical and H2O2 were comparatively decreased in curcumin groups by evaluation of ethidium and DCF fluorescence. According to the results, curcumin can improve sperm kinetic variables and alleviate ROS induced cryoinjury to pig sperm.

• 한국수정란이식학회지
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• 제28권1호
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• pp.31-39
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• 2013

Alpha-tocopherol as an antioxidant acts in preservation of chilled semen by preserving cell membrane damage from lipid peroxidation. Optimum concentrations of ${\alpha}$-tocopherol in egg yolk-citrate (EYC) extender need to be studied in crossbred bull's semen. Different concentrations of ${\alpha}$-tocopherol viz. 0, 1, 2, 4 and 6mg per ml of extender were used. Semen was collected once a week from four bulls used to regular collection, aged 4 to 7 years, weighing 320 to 450 kg, and with body condition score 4 to 4.5 and scrotal circumference 23 to 32 cm. Semen was evaluated routinely and sperm morphology was viewed under light microscope at ${\times}1,000$ magnification after fixing with buffered formal saline. Over 90% had normal head, acrosome, mid-piece and tail. Semen was diluted with egg-yolk-citrate extender to produce $15{\times}10^6$ spermatozoa/ml and 0, 1, 2, 4 and 6 mg/ml ${\alpha}$-tocopherol were added. The semens amples were kept at $8^{\circ}C$. Sperm motility and viability were examined daily up to 5 days under light microscopy at ${\times}200$ magnification. Sperm viability was acceptable (${\geq}40%$) up to the $4^{th}$ day with all concentrations of ${\alpha}$-tocopherol and up to the $5^{th}$ day with 2 mg/ml ${\alpha}$-tocopherol. Sperm motility was acceptable (${\geq}40%$) up to the $3^{rd}$ day irrespective of ${\alpha}$-tocopherol concentration, and up to the $4^{th}$ day with 2 mg/ml ${\alpha}$-tocopherol. It is suggested that the lifespan of chilled semen may be extended up to 4 days by adding 2mg/ml ${\alpha}$-tocopherol.

• Reproductive and Developmental Biology
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• 제41권4호
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• pp.65-70
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• 2017

Alpha-linolenic acid (ALA) is one of n-3 polyunsaturated fatty acids and found mainly in the chloroplasts. Many studies have been reported that intracellular reactive oxygen species (ROS) in mammalian oocytes were reduced by supplementation of ALA in in vitro maturation (IVM) medium. Based on these reports, we expected that ALA acts as an antioxidant during IVM of porcine oocytes. Therefore, the objective of this study was to investigate the antioxidant effect of ALA supplementation during IVM in porcine oocytes. The cumulus-oocyte complexes (COCs) were incubated in IVM medium containing $200{\mu}m$ $H_2O_2$ or $H_2O_2$ with $50{\mu}m$ ALA for 44 h. Nuclear maturation stage of oocytes was evaluated using aceto-orcein method. For measurement of oxidative stress state, intracellular ROS and glutathione (GSH) levels were measured using carboxy-DCFDA and cell tracker red, respectively. In results, oocytes in metaphase-II (MII) stage development was significantly reduced in $H_2O_2$ group compared to non-treated control group $61.84{\pm}1.42%$ and 80.00%, respectively; p<0.05) and it was slightly recovered by treatment of ALA ($69.76{\pm}1.67%$; p<0.05). The intracellular GSH levels was decreased in $H_2O_2$ groups compared with control groups, but it was enhanced by ALA treatment (p<0.05). On the contrary, $H_2O_2$ treatment increased intracellular ROS level in oocytes and $H_2O_2$-induced ROS was decreased by treatment of ALA (p<0.05). Our findings suggested that ALA treatment under oxidative stress condition improve oocyte maturation via elevated GSH and reduced ROS levels in oocytes. Therefore, these results suggest that ALA have an antioxidative ability and it could be used as antioxidant in in vitro production system of porcine embryo.

• 한국식품영양과학회지
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• 제36권4호
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• pp.499-502
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• 2007

본 연구에서는 쌀 섭취의 중요성에 착안하여 건강 기능성 쌀로 개발된 특수미의 항산화 성분을 분석하고자 하였다. 특수미의 polyphenol, phytic acid, anthocyanin은 비색법으로 vitamin E는 순상 HPLC로 분석하였다. 본 연구 결과 일반백미에 비해 특수미는 그 종류에 따라 약 $2{\sim}9$배의 높은 total polyphenol 함유하는 것으로 확인되었다. 또한 쌀에는 insoluble polyphenol이 total polyphenol의 약 $25{\sim}47%$를 차지하는 것으로 나타났다. 쌀의 vitamin E 함량은 백미(0.23mg ${\alpha}-TE/100g$)에 비해 현미(1.02mg ${\alpha}-TE/100g$) 및 특수미 $(0.97{\sim}1.21mg\;{\alpha}-TE/100 g)$에서 높은 함량을 보였고 phytic acid 함량은 백미, 현미, 거대배아미, 녹미, 적색미, 고아미, 흑미 순으로 각각 529, 1,429, 1,507, 1,561, 1,732, 2,163mg/100g으로 나타났다. 일반미와 특수미의 anthocyanin 함량을 측정한 결과 흑미 (302mg/100g)를 제외한 나머지 시료에서는 검출되지 않았으며 일반미와 특수미의 anthocyanin 함량 비교를 위해서는 HPLC 등을 이용한 정밀한 방법이 적용되어야 하겠다. 본 연구 결과는 쌀의 항산화 성분과 항산화 활성 연구에 있어 기초 자료로서 활용될 것으로 예상되며, 건강증진 식품으로서의 쌀을 인식시켜 나아가 쌀의 소비 촉진에 상당한 영향을 미칠 것을 기대된다.

• Asian-Australasian Journal of Animal Sciences
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• 제21권3호
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• pp.340-345
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• 2008

Oxidative stress is one of the major causes of failure of in vitro storage of boar semen. Reactive oxygen species (ROS) are one of the important mediators of oxidative stress during in vitro storage of boar semen. Our study examined the effects of taurine on sperm characteristic and on in vitro developmental embryos during in vitro storage of boar semen for 7 days. Semen was randomly aliquoted into 3 centrifuge tubes and treated with different concentrations of taurine (25-100 mM). The characteristics of boar sperm were analyzed for motility by light microscopy, viability by using a Makler counting chamber and membrane integrity by a hypoosmotic swelling test (HOST). The percentages of motile spermatozoa in taurine groups after 5 days were significantly higher compared to the control. Sperm viability in the control was lower than in taurine groups after 7 days irrespective of different taurine concentration. In the hyoosmotic swelling test (HOST), significantly higher results were obtained in taurine groups after 3 days. Also, the developmental rates of IVM/IVF porcine embryos from semen treated with pyruvate and taurine were significantly increased when compared with the control (p<0.05). These results indicate that supplementation of taurine as an antioxidant in boar semen extender can improve the semen quality.

• 동아시아식생활학회지
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• 제14권4호
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• pp.338-342
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• 2004

This study was performed to measure the antioxidant effects of red ginseng extracts which antioxidation had been promoted through enzyme hydrolysis. In order to observe their tumor-suppressing effects, an anti-cancer medicine and Saponin-SOD, which was a highly antioxidant beverage made from red ginseng saponin adding SOD-like rice (with embryo buds) extracts, were administered to nude mice with large intestine cancer induced. There was a significant increase in the content of phenolic compounds as the enzyme was added. The red ginseng extracts showed a high electron-donating ability with the passage of time. The electron-donating ability was particularly high in the enzyme-treated red ginseng extract, and also observed as high in Saponin-SOD. The lipid-peroxide generation was inhibited depending on the concentration of Saponin-SOD added; the addition of 0.625% Saponin-SOD served to decrease the inhibition level up to 65% compared with the case of no addition (100%). As a result, it could be assumed that Saponin-SOD would strongly inhibit the oxidation of ghost membrane. After the cancer was induced in nude mice through the injection of SNUC-4 cell, there was a significant inhibition in the growth of tumors in nude mice into which Saponin-SOD were injected; the growth of tumors was gradually decreasing with the passage of time after the cancer induction. In particular, when Saponin-SOD was administered together with an anti-cancer medicine, the synergic effect was observed. In conclusion, Saponin-SOD, when used with an anti-cancer medicine, is expected to reduce the amount of free radical and lipid peroxide, which are known to cause harmful effects occurring from the internal application of medicine.

• Environmental Analysis Health and Toxicology
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• 제26권
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• pp.6.1-6.8
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• 2011

Objectives: The present study investigated the potential adverse effects of multi-wall carbon nanotubes (MWCNTs) on pregnant dams and embryonic development following maternal exposure in rats. Methods: MWCNTs were orally administered to pregnant rats from gestational day (GD) 6 through 19 at dose levels of 0, 8, 40, 200, and 1000 mg/kg/day. During the test period, clinical signs, mortality, body weights, food consumption, serum biochemistry, oxidant-antioxidant status, gross findings, organ weights, and Caesarean section findings were examined. Results: All animals survived to the end of the study. A decrease in thymus weight was observed in the highest dose group. However, maternal body weight, food consumption, serum biochemical parameters, and oxidant-antioxidant balance in the kidneys were not affected by treatment with MWCNTs. No treatment-related differences in gestational index, embryo-fetal mortality, or fetal and placental weights were observed between treated and control groups. Conclusions: The results show that 14-day repeated oral dosing of MWCNTs during pregnancy induces minimal maternal toxicity at 1000 mg/kg/day in rats. Under these experimental conditions, the no-observed-adverse-effect level of MWCNTs is considered to be 200 mg/kg/day for dams and 1000 mg/kg/day for embryonic development.

• Journal of Microbiology and Biotechnology
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• 제29권8호
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• pp.1193-1203
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• 2019

We investigated the protective effects of 3-bromo-4,5-dihydroxybenzaldehyde (BDB) from Polysiphonia morrowii Harvey against hydrogen peroxide ($H_2O_2$)-induced apoptosis in Vero cells. BDB exhibited scavenging activity for DPPH, hydroxyl, and alkyl radicals. BDB also inhibited $H_2O_2$-induced lipid peroxidation, cell death, and apoptosis in Vero cells by inhibiting the production of ROS. To evaluate the molecular mechanisms of apoptosis inhibition, the expression of Bax/Bcl-xL and $NF-{\kappa}B$ was assessed by western blot assay. BDB significantly suppressed the cleavage of caspase-9 and PARP and reduced Bax levels in $H_2O_2$-induced Vero cells. Besides, BDB suppressed the phosphorylation of $NF-{\kappa}$B and the translocation of p65 in $H_2O_2$-induced cells. Furthermore, we evaluated the effect of BDB on ROS production, cell death, and lipid peroxidation in an $H_2O_2$-stimulated zebrafish embryo model. Taken together, these results indicated that ROS generation and cell death were significantly inhibited by BDB in zebrafish embryos, thereby proving that BDB exerts excellent antioxidant activity in vitro and in vivo.