• Animal Bioscience
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• 제34권6호
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• pp.975-984
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• 2021

Objective: Inbreeding depression of reproduction is a major concern in the conservation of native chicken genetic resources. Here, based on the successful development of strongly inbred (Sinb) and weakly inbred (Winb) Langshan chickens, we aimed to evaluate inbreeding effects on reproductive traits and identify candidate genes involved in inbreeding depression of reproduction in Langshan chickens. Methods: A two-sample t-test was performed to estimate the differences in phenotypic values of reproductive traits between Sinb and Winb chicken groups. Three healthy chickens with reproductive trait values around the group mean values were selected from each of the groups. Differences in ovarian and hypothalamus transcriptomes between the two groups of chickens were analyzed by RNA sequencing (RNA-Seq). Results: The Sinb chicken group showed an obvious inbreeding depression in reproduction, especially for traits of age at the first egg and egg number at 300 days (p<0.01). Furthermore, 68 and 618 differentially expressed genes (DEGs) were obtained in the hypothalamus and ovary between the two chicken groups, respectively. In the hypothalamus, DEGs were mainly enriched in the pathways related to vitamin metabolism, signal transduction and development of the reproductive system, such as the riboflavin metabolism, Wnt signaling pathway, extracellular matrix-receptor interaction and focal adhesion pathways, including stimulated by retinoic acid 6, serpin family F member 1, secreted frizzled related protein 2, Wnt family member 6, and frizzled class receptor 4 genes. In the ovary, DEGs were significantly enriched in pathways associated with basic metabolism, including amino acid metabolism, oxidative phosphorylation, and glycosaminoglycan degradation. A series of key DEGs involved in folate biosynthesis (gamma-glutamyl hydrolase, guanosine triphosphate cyclohydrolase 1), oocyte meiosis and ovarian function (cytoplasmic polyadenylation element binding protein 1, structural maintenance of chromosomes 1B, and speedy/RINGO cell cycle regulator family member A), spermatogenesis and male fertility (prostaglandin D2 synthase 21 kDa), Mov10 RISC complex RNA helicase like 1, and deuterosome assembly protein 1) were identified, and these may play important roles in inbreeding depression in reproduction. Conclusion: The results improve our understanding of the regulatory mechanisms underlying inbreeding depression in chicken reproduction and provide a theoretical basis for the conservation of species resources.

• 한국응용과학기술학회지
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• 제35권4호
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• pp.1250-1259
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• 2018

여주(Momordica charantia) 열수 추출물을 젖산균(Leuconostoc mesenteroidesies)으로 발효하여 얻은 생산물의 항산화 및 항당뇨 효과를 관찰하였다. 여주 발효물(MC-LM)과 여주 추출물(MC)의 항산화 활성은 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)(ABTS) radical 소거 측정법을 사용하여 관찰하였다. MC 처리군은 radical 소거활성이 거의 나타나지 않았으나 MC-LM의 경우 $500{\mu}g/mL$ 농도에서 47%의 활성이 관찰되었으며 MC-LM의 농도가 증가함에 따라 활성이 유의하게(p<0.05) 증가하는 것을 확인 할 수 있었다. MC-LM은 MC에 비해 peroxyl radical에 의한 DNA의 산화적 손상을 더 효과적으로 억제하였으며 MC-LM의 농도가 증가함에 따라 strand breakage의 억제가 유의적으로 증가 하였다(p<0.05). 혈당량과 밀접한 관련이 있는 ${\alpha}-glucosidase$의 활성 억제정도를 측정한 결과 MC는 8 mg/mL 농도에서 ${\alpha}-glucosidase$의 활성을 30.1% 감소시켰고 같은 농도에서 MC-LM은 58.9% 감소 시켰다. 또한 alloxan에 의한 RIN-m5F 췌장세포 사멸에 발효물의 영향을 관찰한 결과 MC-LM을 농도별로 처리한 군에서 세포생존율이 유의적으로(p<0.05) 증가하였으며 $1000{\mu}g/mL$ 농도에서 MC군 보다 20% 높게 증가되었다. Alloxan에 의한 ${\beta}-cell$파괴를 유도하여 RIN-m5F cell이 분비한 인슐린의 측정결과 MC-LM을 농도별로 첨가한 모든 군에서 인슐린 분비능이 증가하는 경향을 보였으며, $1000{\mu}g/mL$ 농도에서 MC군 보다 15% 높게 증가되었다. 결론적으로 본 연구에서 관찰된 결과들을 통해 여주 발효물은 항산화 및 항당뇨 효과가 있는 것으로 확인되었다.

• 생명과학회지
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• 제28권12호
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• pp.1469-1476
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• 2018

Warburg 효과의 발생은 고형암에서 화학적 항암제의 내성을 발생시킨다. 따라서 호기성 해당과정과 같은 에너지 대사과정은 암 치료의 중요한 표적으로 알려져 있다. Pyruvate dehydrogenase kinase (PDK) 활성 억제물질로 알려진 dichloroacetate (DCA)는 많은 암세포에서 포도당의 호기성 해당과정을 산화적인산화 과정으로 전환시킬 수 있음이 보고되었다. 이 연구는 치료가 매우 어렵다고 알려진 인간 역분화 갑상선 암세포주인 8505C의 성장에 미치는 DCA효과를 조사하였다. DCA는 정상 갑상선 세포주의 성장에는 영향을 주지 않은 반면 8505C 세포주의 성장을 특이적으로 저해하였다. DCA의 처리에 의해 8505C 세포주는 $HIF1{\alpha}$, PDK1, Bcl-2와 같은 항-세포자살 관련 단백질들의 발현이 감소하고, Bax와 p21과 같은 세포자살 유도 단백질과 세포주기 억제 단백질의 증가로 인하여 세포주기 정지와 세포자살 유도에 의해 성장이 억제되었다. 이런 세포의 변화는 DCA 처리에 의한 활성산소족의 생산이 증가하고, 포도당 대사가 호기성 해당과정에서 산화적인산화 과정으로 전환되었기 때문이란 것을 확인하였다. 흥미롭게도, DCA는 포도당 대사과정의 변화뿐만 아니라 sodium/iodine symporter (NIS)의 발현양도 증가시킨다는 것을 확인하였다. 이 연구의 결과로 PDK 활성 저해제는 치료하기 힘든 역분화 갑상선 암 치료제로 이용할 수 있고, 또한 역분화 갑상선 암에 대한 방사능 치료의 효능을 높일 수 있을 것으로 기대된다.

• 생명과학회지
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• 제29권9호
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• pp.977-985
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• 2019

쑥부쟁이는 국화과에 속하는 다년생 식물로서 다양한 질병의 예방 및 치료에 오랫동안 사용되어 왔다. 최근 연구에서 쑥부쟁이 잎 추출물이 항산화 및 항염증 효과가 있는 것으로 알려져 있지만, 정확한 효능 평가에 관한 연구는 여전히 미비한 실정이다. 본 연구에서는 RAW 264.7 대식세포에서 쑥부쟁이 잎 에탄올 추출물(EEAY)의 항산화 효능이 항염증 효능과 연관이 있는지의 여부를 조사하였다. 본 연구의 결과에 의하면, EEAY는 $H_2O_2$ 처리에 의한 RAW 264.7 세포의 세포 독성을 유의적으로 억제시켰으며, 이는 Nrf2 및 HO-1의 발현 증가와 관련이 있음을 보여 주었다. 또한 EEAY는 $H_2O_2$에 의한 apoptosis를 유의적으로 억제하였으며, 이는 caspase-3의 활성 억제에 따른 PARP의 분해 차단과 연관성이 있었다. 그리고 EEAY는 대표적인 항 염증성 사이토카인인 IL-10의 발현 및 생산을 증가시켰으며, 이는 전사 및 번역 수준에서의 TLR-4 및 Myd88 발현 증가와 관련이 있었다. 아울러 EEAY는 LPS에 의한 염증성 매개인자인 NO의 생성 증가를 현저히 억제하였으며, EEAY에 의한 NO 생성의 억제 효과는 HO-1 유도제인 hemin에 의해 더욱 증가되었다. 따라서 본 연구의 결과는 EEAY에 의한 산화적 및 염증성 스트레스에 대한 RAW 264.7 대식세포의 보호 효과에 최소한 Nrf2/HO-1 신호 경로의 활성화가 관여할 가능성을 보여주었다.

• 한국동물생명공학회지
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• 제34권1호
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• pp.10-19
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• 2019

Morphology of cumulus-oocyte-complexes (COCs) at germinal vesicle (GV) stage as one of the evaluation criteria for oocyte maturation quality after in vitro maturation (IVM) plays important roles on the meiotic maturation, fertilization and early embryonic development in pigs. When cumulus cells of COCs are insufficient, which is induced the low oocyte maturation rate by the increasing of reactive oxygen species (ROS) in porcine oocyte during IVM. The ROS are known to generate including superoxide and hydrogen peroxide from electron transport system of mitochondria during oocyte maturation in pigs. To regulate the ROS production, the cumulus cells is secreted the various antioxidant enzymes during IVM of porcine oocyte. Our previous study showed that Mito-TEMPO, superoxide specific scavenger, improves the embryonic developmental competence and blastocyst formation rate by regulating of mitochondria functions in pigs. However, the effects of Mito-TEMPO as a superoxide scavenger to help the anti-oxidant functions from cumulus cells of COCs on meiotic maturation during porcine oocyte IVM has not been reported. Here, we categorized experimental groups into two groups (Grade 1: G1; high cumulus cells and Grade 2: G2; low cumulus cells) by using hemocytometer. The meiotic maturation rate from G2 was significantly (p < 0.05) decreased (G1: $79.9{\pm}3.8%$ vs G2: $57.5{\pm}4.6%$) compared to G1. To investigate the production of mitochondria derived superoxide, we used the mitochondrial superoxide dye, Mito-SOX. Red fluorescence of Mito-SOX detected superoxide was significantly (p < 0.05) increased in COCs of G2 compared with G1. And, we examined expression levels of genes associated with mitochondrial antioxidant such as SOD1, SOD2 and PRDX3 using a RT-PCR in porcine COCs at 44 h of IVM. The mRNA levels of three antioxidant enzymes expression in COCs from G2 were significantly (p < 0.05) lower than COCs of G1. In addition, we investigated the anti-oxidative effects of Mito-TEMPO on meiotic maturation of porcine oocyte from G1 and G2. Meiotic maturation and mRNA levels of antioxidant enzymes were significantly (p < 0.05) recovered in G2 by Mito-TEMPO ($0.1{\mu}M$, MT) treatment (G2: $68.4{\pm}3.2%$ vs G2 + MT: $73.9{\pm}1.4%$). Therefore, our results suggest that reduction of mitochondria derived superoxide by Mito-TEMPO may improves the meiotic maturation in IVM of porcine oocyte.

• 한국수정란이식학회지
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• 제33권4호
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• pp.211-220
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• 2018

Nitric oxide (NO) has an important role in oocyte maturation and embryonic development in mammals. This study examined the effect of exogenous NO donor S-nitroso-N-acetylpenicillamine (SNAP) in a maturation medium on meiotic progression and embryonic development after parthenogenesis (PA) and somatic cell nuclear transfer (SCNT) in pigs. When oocytes were exposed to $0.1{\mu}M$ SNAP for first 22 h of in vitro maturation (IVM) in Experiment 1, SNAP significantly improved blastocyst development in both defined and standard follicular fluid-supplemented media compared to untreated control (48.4 vs. 31.7-42.5%). SNAP treatment significantly arrested meiotic progression of oocytes at the germinal vesicle stage at 11 h of IVM (61.2 vs. 38.7%). However, there was no effect on meiotic progression at 22 h of IVM (Experiment 2). In Experiment 3, when oocytes were treated with SNAP at 0.001, 0.1 and $10{\mu}M$ during the first 22 h of IVM to determine a suitable concentration, $0.1{\mu}M$ SNAP (54.2%) exhibited a higher blastocyst formation than 0 and $10{\mu}M$ SNAP (36.6 and 36.6%, respectively). Time-dependent effect of SNAP treatment was evaluated in Experiment 4. It was observed that SNAP treatment for the first 22 h of IVM significantly increased blastocyst formation compared to no treatment (57.1% vs. 46.2%). Antioxidant effect of SNAP was compared with that of cysteine. SNAP treatment significantly improved embryonic development to the blastocyst stage (49.1-51.5% vs. 34.4-37.5%) irrespective of the presence or absence of cysteine (Experiment 5). Moreover, SNAP significantly increased glutathione (GSH) content and inversely decreased the reactive oxygen species (ROS) level and mitochondrial oxidative activity in IVM oocytes. SNAP treatment during IVM showed a stimulating effect on in vitro development of SCNT embryos (Experiment 7). These results demonstrates that SNAP improves developmental competence of PA and SCNT embryos probably by maintaining the redox homeostasis through increasing GSH content and mitochondrial quality and decreasing ROS in IVM oocytes.

• 생명과학회지
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• 제29권2호
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• pp.191-197
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• 2019

파킨슨병은 운동완서, 근육경직, 진전 및 비정상적인 자세 등을 임상적 특징으로 하는 주로 운동 신경계에 영향을 주는 진행성 신경 퇴행성 질환이다. 파킨슨병은 산화 스트레스와 세포 내 신호 전달 경로의 조절 장애에 의한 뇌 흑색치밀부에서의 도파민성 신경세포의 사멸을 특징으로 한다. Quercetin의 주요 대사산물인 Quercetin-3-O-glucuronide (Q3GA)는 신경 보호 효과가 있는 것으로 보고 되어 왔다. 본 연구에서는 SH-SY5Y 세포에서 1-methyl-4-phenyl pyridinium ($MPP^+$)에 의해 유도된 신경 독성에 대한 Q3GA의 신경 보호 효과와 그 분자 조절 기전을 조사하였다. Q3GA는 $MPP^+$에 의해 유도된 세포 사멸을 유의적으로 감소시켰으며 PARP 절단을 감소시켰다. 또한, Bax/Bcl-2 비율의 감소와 함께 $MPP^+$에 의해 증가된 세포 내 ROS를 감소시켰다. Q3GA는 $MPP^+$에 의해 감소된 Akt와 CREB의 인산화를 유의적으로 회복시켰지만, ERK에는 영향을 미치지 않았다. 이 결과는 Q3GA가 ROS 생산 억제와 Akt/CREB 신호 전달 경로를 통해 $MPP^+$ 에 의해 유도된 신경 독성을 억제시킬 수 있음을 시사한다. 본 연구는 Q3GA가 파킨슨병에 대한 예방제 또는 치료제로 개발될 수 있는 가능성을 제시한다.

• Asian-Australasian Journal of Animal Sciences
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• 제32권8호
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• pp.1112-1121
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• 2019

Objective: Gram-negative bacteria lipopolysaccharide (LPS) has been reported to be associated with uterine impairment, embryonic resorption, ovarian dysfunction, and follicle retardation. Here, we aimed to investigate the toxic effects of LPS on the maturation ability and parthenogenetic developmental competence of bovine oocytes. Methods: First, we developed an in vitro model to study the response of bovine cumulusoocyte complexes (COCs) to LPS stress. After incubating germinal vesicle COCs in $10{\mu}g/mL$ of LPS, we analyzed the following three aspects: the expression levels of the LPS receptor toll-like receptor 4 (TLR4) in COCs, activities of intracellular signaling protein p38 mitogen-activated protein kinase (p38 MAPK) and nuclear factor-kappa B (NF-${\kappa}B$); and the concentrations of interleukin (IL)-$1{\beta}$, tumor necrosis factor (TNF)-${\alpha}$, and IL-6. Furthermore, we determined the effects of LPS on the maturation ability and parthenogenetic developmental competence of bovine oocytes. Results: The results revealed that LPS treatment significantly elevated TLR4 mRNA and protein expression levels in COCs. Exposure of COCs to LPS also resulted in a marked increase in activity of the intracellular signaling protein p-p38 MAPK and NF-${\kappa}B$. Furthermore, oocytes cultured in maturation medium containing LPS had significantly higher concentrations of the proinflammatory cytokines IL-$1{\beta}$, TNF-${\alpha}$, and IL-6. LPS exposure significantly decreased the first polar body extrusion rate. The cytoplasmic maturation, characterized by polar body extrusion and distribution of peripheral cortical granules, was significantly impaired in LPS-treated oocytes. Moreover, LPS exposure significantly increased intracellular reactive oxygen species levels and the relative mRNA abundance of the antioxidants thioredoxin (Trx), Trx2, and peroxiredoxin 1 in oocytes. Moreover, the early apoptotic rate and the release of cytochrome C were significantly increased in response to LPS. The cleavage, morula, and blastocyst formation rates were significantly lower in parthenogenetically activated oocytes exposed to LPS, while the incidence of apoptotic nuclei in blastocysts was significantly increased. Conclusion: Together, these results provide an underlying mechanism by which LPS impairs maturation potential in bovine oocytes.

• Fisheries and Aquatic Sciences
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• 제24권1호
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• pp.19-31
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• 2021

Seaweeds are a potential source of minerals, essential amino acids, fatty acids, proteins, and various bioactive compounds such as antioxidants. The higher water content of seaweeds reduces the shelf life and this requires the appropriate drying method. The drying conditions play a major role in the conservation of nutrient composition in dried seaweeds. In recent years, the seaweed industry has used many different drying methods with advantages and limitations. Hybrid hot-water Goodle dryer (HHGD) which is a special dryer mixed with hot-water and a Korean traditional heating system (Goodlejang) might be a solution to avoid these limitations. The present study evaluated the effect of drying conditions in HHGD on nutrient composition and bioactivities of brown seaweeds. Moreover, freeze-dryer (FD) and HHGD were employed in this study to compare the dried outputs obtained from four brown seaweed species. The present study aims to evaluate the effect of the hybrid hot-water Goodle drying method (HHGDM) on the nutritional composition and antioxidant activity of dried seaweeds. AOAC standard methods were used to analyze the proximate composition of dried samples and their 70% ethanol extract. The intracellular and extracellular antioxidant activities were evaluated using Vero cells and electron spin resonance (ESR) spectrometer respectively. High performance liquid chromatography, apoptotic body formation, and in-vivo experiments were used for further confirmation of the quality of dried output. The proximate composition results obtained from drying in HHGD and FD did not exhibit any significant difference. Moreover, the seaweed extracts from the dried seaweeds by HHGD and FD dryings were also not different and both significantly down-regulated in-vivo and in-vitro oxidative stress. Furthermore, the high performance liquid chromatography results revealed that the two dryers did not make the major peaks different in the chromatograms. Freeze-drying method (FDM) provides elevated quality for dried output, but there are limitations such as high cost and low capacity. The results from a novel HHGD did not provide any significant difference with the results in FD and expressed a potential to avoid the limitations in FD. Overall, these findings solidified the applicability of HHGD over FD.

• 한방비만학회지
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• 제21권1호
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• pp.10-21
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• 2021

Objectives: Benign prostatic hyperplasia (BPH) is a progressive pathological condition characterized by excessive proliferation of the prostate. In this study, we evaluated the effect of Corni Fructus water extract (CF) on the promotion of prostate cell proliferation by dihydrotestosterone (DHT). Methods: The effect of CF on the proliferation of LNCaP prostate cells was evaluated, and DHT was treated to induce an in vitro BPH model. To study the mechanism of inhibition of cell proliferation and BPH by CF, changes in the expression of key factors related to cell cycle and BPH were investigated. We further investigated the effect on the production of reactive oxygen species (ROS) and nitric oxide (NO) to evaluate the antioxidant and anti-inflammatory efficacy of CF. Results: Inhibition of LNCaP cell proliferation by CF was associated with decreased expression of cyclin D1 and cyclin A and increased expression of cyclin-dependent kinase inhibitor p21. CF also suppressed expression of BPH inducing factors such as 5α-reductase type 2 and androgen receptor (AR) as well as prostate specific antigen (PSA). Furthermore, CF significantly blocked DHT-induced LNCaP cell proliferation and effectively attenuated DHT-induced expression of BPH mediators and cyclins. In addition, CF inhibited DHT-induced oxidative and inflammatory reactions by inhibiting production of ROS and NO. Conclusion: Our results demonstrated that CF probably acted as 5α-reductase type 2 inhibitor, preventing the 5α-reductase type 2-AR signaling pathway, thereby reducing the conversion of testosterone to DHT and the expression of PSA, which is at least correlated with the antioxidant and anti-inflammatory activities of CF.