Melatonin Reduces the Bisphenol A-induced Superoxide and Improves Oocyte Maturation through Reducing of Mitochondrial Derived Apoptosis in Pigs

Hyo-Jin Park,Jin-Woo Kim,Seul-Gi Yang1,Min-Ji Kim,Ho-Guen Jegal,Joung Jun Park,Deog-Bon Koo 한국동물생명공학회(구 한국동물번식학회) 2018 발생공학 국제심포지엄 및 학술대회 Vol.2018 No.06

Bisphenol-A (BPA) as an organic synthetic compound of exhibiting estrogen-mimicking and hormone-like properties, which is commonly used to induce cellular stress or female reproductive toxicity. In addition, BPA induces the increasing of mitochondrial derived reactive oxygen species (ROS) such as superoxide, and production of these ROS affects to the meiotic maturation and cumulus cells expansion on in vitro maturation (IVM) of porcine cumulus-oocyte complexes (COCs). However, anti-oxidative effect of melatonin for reduction of BPA-induced superoxide on porcine oocyte maturation has not been reported. Therefore, in present study, we confirmed that the reduction of BPA-derived superoxide by melatonin related to the reducing of mitochondria mediated apoptosis on meiotic maturation and cumulus cells expansion of porcine COCs. Then, to investigate the effects of superoxide specific scavenger, Mito-TEMPO, during porcine oocyte maturation progression, COCs cultured in maturation medium with Mito-TEMPO (0.1 μM) after pre-treatment of BPA (75 μM) for 22 h. Reduced meiotic maturation rate and cumulus cells expansion of COCs in the BPA (75 μM) treated group were recovered (p<0.01) by Mito-TEMPO treatment. Also, increasing of mitochondria derived apoptotic factors (AIF, Cleaved Caspase 3 and Cleaved PARP 1) protein levels by BPA treatment were reduced by Mito-TEMPO treatment in porcine COCs maturation. Positive effects of Mito-TEMPO for superoxide reduction on oocyte maturation and reducing mitochondrial apoptosis showed the same pattern in melatonin (0.1 μM) treated COCs. In case of supplemented with BPA and melatonin, superoxide production in COCs was not changed compared to control or melatonin treated groups. Based on these results, we concluded that melatonin as a regulator of superoxide such as Mito-TEMPO improves oocyte maturation through reduction of mitochondria derived apoptosis during IVM of porcine COCs.

An Exploratory Research on PCC Application of Crystalline Limestone: Effects of Limestone Crystallographic Characteristicson Hydraulic Activity

Yang, Ye-Jin,Jegal, Yu-Jin,Nam, Seong-Young,Kim, Jin,Ahn, Ji-Whan The Korean Ceramic Society 2014 한국세라믹학회지 Vol.51 No.2

Quicklime(CaO) is generally obtained through the calcination of limestone, the main component of which is calcium carbonate($CaCO_3$). Quicklime generates high-temperature heat when reacting with water, forming slaked lime($Ca(OH)_2$). The industrial sectors for limestone are determined by the hydraulic activity of slaked lime, which is obtained by measuring temperature changes during the hydration reaction. Accordingly, this study examined the different crystallographic characteristics of limestone as affected by the geological origins of the regions where the limestones were produced, and how these characteristics affected hydraulic activity. Six limestone samples were collected from the Jecheon and Cheongsong areas and the hydraulic activities were measured in accordance with KS E 3077. The results indicate that limestone produced in the Cheongsong area, recrystallized through metamorphism caused by hydrothermal alteration, hada larger grain size of calcite than that of the Jecheon area, and displays a tendency of changing to marble. Limestone from the Cheonsong area showed more radical reaction in the early stage of hydration compared to that ofthe Jecheon area. In addition, it was revealed that limestone having more impurities like $SiO_2$ have lower hydraulic activity.

동종골수이식 후 폐합병증

제갈양진 ( Jegal Yang Jin ),이제환 ( Lee Je Hwan ),이규형 ( Lee Gyu Hyeong ),김우건 ( Kim U Geon ),심태선 ( Sim Tae Seon ),임채만 ( Im Chae Man ),고윤석 ( Go Yun Seog ),이상도 ( Lee Sang Do ),김우성 ( Kim U Seong ),김원동 ( Kim Won 대한결핵 및 호흡기학회 2000 Tuberculosis and Respiratory Diseases Vol.49 No.2

Exogenous Ganglioside GT1b Enhances Oocytes Maturation, Cumulus Cells Expansion and Further Preimplantation Developmental Competence in Pigs

Jin-Woo Kim,Hyo-Jin Park,Seul-Gi Yang,Jae-Min Jung,Min-Ji Kim,In-Su Kim,Ho-Guen Jegal,Deog-Bon Koo 한국수정란이식학회 2017 한국수정란이식학회 학술대회 Vol.2017 No.05

Ganglioside GT1b, glycosphigolipids with three sialic acid, is known to play an important role in signal transduction such as epidermal growth factor receptor (EGFR). EGF is also known to induce resumption of meiosis and cumulus cells expansion during porcine oocyte maturation. Therefore, this study was conducted to evaluate the effects of ganglioside GT1b on resumption of meiosis and cumulus cells expansion in porcine oocyte maturation. First, porcine cumulus-oocyte complexes were cultured in NCSU-23 medium supplemented with GT1b (0, 1, 2 and 4 μM) at 44 h. We observed that the proportion of the metaphase II (M II) stage was significantly increased in the 2 μM GT1b (78.0 ± 2.3) treated group than in the other groups. Furthermore, expression of cumulus cells expansion factor genes (Has2, TNFAIP6, Ptx3) were significantly increased in the 2 μM GT1b treated group than in the other groups. Next, we investigated the meiotic maturation and the expressions of cumulus cells expansion factor genes after GT1b and/or EGF treatment. The proportion of the M II stage was significantly higher in the GT1b+EGF (90.1 ± 2.3) treated group than in the other groups. Moreover, expressions of cumulus cells expansion factor genes were significantly increased in the GT1b+EGF treated group than in the control group. After in vitro fertilization, fertilization rate, preimplantation development competence and quality of blastocyst were improved in oocytes derived from GT1b+EGF treated group. Taken together, these results suggest that exogenous ganglioside GT1b improving the developmental competence of porcine embryos via increase of resumption of meiosis and cumulus cells expansion during in vitro maturation of porcine oocytes.

Reversible Effects of Exogenous GM3 on Meiotic Maturation and Cumulus Cells Expansion of Porcine Cumulus-oocyte Complexes

Jin-Woo Kim,Hyo-Jin Park,Jae-Min Jung,Seul-Gi Yang,Min-Ji Kim,In-Su Kim,Ho-Geun Jegal,Deog-Bon Koo 한국수정란이식학회 2018 한국동물생명공학회지 Vol.33 No.4

Ganglioside GM3 is known as an inhibition factor of cell differentiation and proliferation via inhibition of epidermal growth factor receptor (EGFR) phosphorylation. Our previous study showed that the exogenous ganglioside GM3 reduced the meiotic maturation of porcine oocytes and induced apoptosis at 44 h of in vitro maturation (IVM). However, the role of ganglioside GM3 in the relationship between EGFR signaling and apoptosis during porcine oocyte maturation has not yet been studied. First, porcine cumulus-oocyte complexes (COCs) were cultured in the NCSU-23 medium with exogenous ganglioside GM3 according to maturation periods (non-treated, only IVM I: 0 - 22 h, only IVM II: 22 - 44 h and IVM I & II: 0 - 44 h). We confirmed that the proportion of germinal vesicle breakdown (GVBD) increased significantly in the IVM I treated group than in the control group. We also confirmed that the meiotic maturation until M II stage and polar body formation decreased significantly in the only IVM I treated group. Cumulus cell expansion and mRNA levels of the expansion-related factors (HAS2, TNFAIP6 and PTX3) decreased significantly in the IVM I treated group than in the control group. Protein levels of EGFR, p-EGFR, ERK1/2, and p-ERK1/2 decreased significantly in the GM3-treated groups, during the IVM I period. In addition, cellular apoptosis, determined using TUNEL assay, and protein levels of Cleaved caspase 3, were increased significantly in the GM3-treated COCs during the IVM I period. Based on these results, ganglioside GM3 exposure of porcine COCs during the IVM I period reduced meiotic maturation and cumulus cell expansion via inhibition of EGFR activity in pigs.

Reversible Effects of Exogenous GM3 on Meiotic Maturation and Cumulus Cells Expansion of Porcine Cumulus-oocyte Complexes

Kim, Jin-Woo,Park, Hyo-Jin,Jung, Jae-Min,Yang, Seul-Gi,Kim, Min-Ji,Kim, In-Su,Jegal, Ho-Geun,Koo, Deog-Bon The Korean Society of Embryo Transfer 2018 한국동물생명공학회지 Vol.33 No.4

Ganglioside GM3 is known as an inhibition factor of cell differentiation and proliferation via inhibition of epidermal growth factor receptor (EGFR) phosphorylation. Our previous study showed that the exogenous ganglioside GM3 reduced the meiotic maturation of porcine oocytes and induced apoptosis at 44 h of in vitro maturation (IVM). However, the role of ganglioside GM3 in the relationship between EGFR signaling and apoptosis during porcine oocyte maturation has not yet been studied. First, porcine cumulus-oocyte complexes (COCs) were cultured in the NCSU-23 medium with exogenous ganglioside GM3 according to maturation periods (non-treated, only IVM I: 0 - 22 h, only IVM II: 22 - 44 h and IVM I & II: 0 - 44 h). We confirmed that the proportion of germinal vesicle breakdown (GVBD) increased significantly in the IVM I treated group than in the control group. We also confirmed that the meiotic maturation until M II stage and polar body formation decreased significantly in the only IVM I treated group. Cumulus cell expansion and mRNA levels of the expansion-related factors (HAS2, TNFAIP6 and PTX3) decreased significantly in the IVM I treated group than in the control group. Protein levels of EGFR, p-EGFR, ERK1/2, and p-ERK1/2 decreased significantly in the GM3-treated groups, during the IVM I period. In addition, cellular apoptosis, determined using TUNEL assay, and protein levels of Cleaved caspase 3, were increased significantly in the GM3-treated COCs during the IVM I period. Based on these results, ganglioside GM3 exposure of porcine COCs during the IVM I period reduced meiotic maturation and cumulus cell expansion via inhibition of EGFR activity in pigs.

파라쿼트 중독 환자에서 deferoxamine의 치료 효과

허진원 ( Jin Won Huh ),제갈양진 ( Yang Jin Jegal ),홍상범 ( Sang Bum Hong ),오연목 ( Yeon Mok Oh ),심태선 ( Tae Sun Shim ),임채만 ( Chae Man Lim ),이상도 ( Sang Do Lee ),김우성 ( Woo Sung Kim ),김동순 ( Dong Soon Kim ),김원동 ( Won 대한결핵 및 호흡기학회 2007 Tuberculosis and Respiratory Diseases Vol.62 No.2

연구배경: 파라쿼트 중독은 다발성 장기부전과 폐섬유화를 유발하여 높은 사망률을 초래한다. 폐섬유화의 약화를 위해 여러 종류의 면역억제제가 사용되었으나 그 치료효과는 다양하다. 철은 파라쿼트로 유발된 산화 스트레스에 의한 폐손상에 중요한 역할은 한다. 본 연구는 파라쿼트 중독의 치료시 철 킬레이트 제제인 deferoxamine의 효과를 보고자 시행되었다. 방법: 2001년 10월부터 2005년 4월까지 파라쿼트 중독으로 내과계중환자실에 입원한 37명 중 28명이 본 연구에 포함되었다. 환자들은 서울아산병원의 파라쿼트 치료 프로토콜에 따라서 치료되었다. 이 중 12명이 추가로 deferoxamine 투여군으로 무작위 분류되었다. 결과: 두 군간의 성별, 나이, 파라쿼트 중독의 정도, 섭취 후 병원까지의 도착 시간 등은 유의한 차이를 보이지 않았다. 파라쿼트 섭취 후 간기능과 신기능의 변화도 유의한 차이를 보이지 않았다. 전체 사망률은 두 군 간의 차이는 없었으나 deferoxamine 투여군에서 대조군과 비교시 호흡부전의 빈도가 높았다(4/7 versus 0/9, p=0.019). 결론: 파라쿼트 중독 시 본 연구에서 적용된 용량과 기간의 deferoxamine (100mg/kg during 24h)은 치료효과가 없을 것으로 사료된다. Background: Paraquat is known to induce oxidant injury that results in multiorgan failure and lung fibrosis. Iron has been considered to play a key role in paraquat-induced oxidant lung injury. This study examined the effect of deferoxamine, an iron-chelating agent, in the treatment of paraquat poisoning. Methods: From September, 2001 to April, 2005, 28 patients with paraquat poisoning who were admitted at a medical intensive care unit of a University-affiliated hospital, were enrolled in this study. Sixteen patients were treated according to the paraquat poisoning treatment guidelines and 12 received an intravenous infusion of deferoxamine in addition to the treatment guidelines. Results: There were no differences between the two groups in terms of age, gender, severity of paraquat poisoning, and the time elapsed from ingestion to presentation at hospital. There was no difference in overall mortality between the two groups but the incidence of respiratory failure in the deferoxamine group was higher than in the conventional group(4/7 versus 0/9, p=0.019). Conclusion: Deferoxamine seems to have no clinical benefit compared with the conventional treatment. (Tuberc Respir Dis 2007; 62: 113-118)

Regulation of Mitochondrial Ca2+ by Ruthenium Red during in vitro Fertilization Improves Mitochondrial Functions and Blastocyst Development in Porcine Embryos

Ho-Guen Jegal,Hyo-Jin Park,Jin-Woo Kim,Seul-Gi Yang,Min-Ji Kim,Joung Jun Park,Deog-Bon Koo 한국동물생명공학회(구 한국동물번식학회) 2018 발생공학 국제심포지엄 및 학술대회 Vol.2018 No.06

Mitochondria have vital functions that regulate Ca2+ homeostasis and ATP production in fertilized mammalian embryos. In addition, regulation of mitochondrial Ca2+ is related to the mitochondrial functions and mitochondria mediated apoptosis. However, early embryonic development according to the change of mitochondrial Ca2+ regulation during IVF of porcine oocyte has not been reported. In this study, we investigated the regulation of mitochondrial Ca2+ by ruthenium red (RR) during IVF affects blastocyst development in porcine embryos. Here, we investigated the changes of mitochondrial Ca2+ by using Rhod-2 staining in fertilized oocytes during IVF progression (0, 3 and 6 h). After IVF, Rhod-2 expression significantly increased in most of fertilized oocytes at 3 and 6 h. Therefore, we treated RR into IVF medium for reduction of mitochondrial Ca2+. As expected, expressions of rhod-2 and mito-sox significantly decreased in RR treated zygotes. To confirm the mitochondrial functions, we performed the JC-1 and ATP determination in porcine zygotes. As a result, MMP and ATP increased in fertilized oocytes after RR treatment. Interestingly, blastocyst development rate was significantly increased in 20 μM RR treated fertilizing oocytes compared to other groups (p <0.05; 20 μM RR: 33.2±5.0% vs. 10 μM RR: 22.4±5.0% and Con: 24.7±3.8%). In addition, we confirmed that apoptosis in porcine blastocysts was reduced according to RR treatment during IVF procedure by using a TUNEL assay. These results demonstrated that positive effects of RR for regulation of mitochondrial Ca2+ during IVF progression improve the mitochondrial functions and blastocyst developmental competence in porcine embryos.

Reduction of mitochondria derived superoxide by Mito-TEMPO improves the porcine oocyte maturation

Seul-Gi Yang,Hyo-Jin Park,Jin-Woo Kim,Min-Ji Kim,Ho-Guen Jegal,In-Su Kim,Min-Young Guk,Sun-Mi Park,Ji-Eun Lee,Deog-Bon Koo,Joung Jun Park 한국수정란이식학회 2018 한국수정란이식학회 학술대회 Vol.2018 No.11

In general, the shape of cumulus-oocyte-complexes (COCs) at germinal vesicle (GV) stage is important roles on meiotic maturation of porcine oocyte during in vitro maturation (IVM). Then, mitochondria produce reactive oxygen species (ROS) such as superoxide from electron transport system during oocyte maturation. ROS levels on oocytes are regulated by various antioxidant enzymes in cumulus cells (CCs). However, the effect of mitochondria derived superoxide production from CCs during IVM of porcine oocyte has not been reported. Firstly, we divided groups according to large number of CCs (Grade 1: G1) and small number of CCs (Grade 2: G2). Then, we counted cumulus cells of G1 and G2 oocyte by using haemocytometer. The oocyte maturation rate was significant decreased (p < 0.05) in G2 oocytes than that of G1 oocytes. We measured mitochondria derived superoxide in G1 and G2 COCs by using Mito-SOX staining. Mitochondrial superoxide was higher in G2 COCs than G1 COCs. Then, the mRNA expression levels of antioxidant enzymes (SOD1, SOD2 and PRDX3) in G2 COCs were decreased compared to G1 COCs. To reduce mitochondria derived superoxide, we used Mito-TEMPO as mitochondrial superoxide scavenger. Oocyte maturation rates in both G1 and G2 groups treated with Mito-TEMPO were increased than that of non-treated groups. Mitochondrial superoxide was lower in G1 and G2 groups treated with Mito-TEMPO than that of non-treatment groups. The mRNA expression levels of antioxidant enzymes in G1 and G2 COCs treated with Mito-TEMPO were increased compared to non-treated groups. Based on these findings, we suggest that reduction of mitochondria derived superoxide by Mito-TEMPO assists maturation competence in porcine oocytes.

Melatonin Improves Oocyte Maturation and Mitochondrial Functions by Reducing Bisphenol A-Derived Superoxide in Porcine Oocytes In Vitro

Park, Hyo-Jin,Park, Soo-Yong,Kim, Jin-Woo,Yang, Seul-Gi,Kim, Min-Ji,Jegal, Ho-Guen,Kim, In-Su,Choo, Young-Kug,Koo, Deog-Bon MDPI 2018 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.19 No.11

<P>Bisphenol A (BPA) is synthetic organic compound that exhibits estrogen-like properties and it induces mitochondrial superoxide production. Melatonin (Mela) protects against BPA-mediated cell damage and apoptosis. However, the antioxidative effects of Mela against BPA-induced superoxide production in porcine oocytes are still not known. In this study, we investigated the antioxidative effects of Mela against BPA-derived superoxide on oocyte maturation in pigs. To investigate the effects of the superoxide specific scavenger, Mito-TEMPO, on porcine oocyte maturation in response to BPA exposure apoptosis proteins, we treated the oocytes with Mito-TEMPO (0.1 µM) after pre-treating them with BPA (75 µM) for 22 h. As expected, the reduction in meiotic maturation and cumulus cell expansion of cumulus-oocyte-complexes (COCs) in the BPA (75 µM) treated group was recovered (<I>p</I> < 0.01) by treatment with Mito-TEMPO (0.1 µM). An increase in the levels of mitochondrial apoptotic proteins (AIF, cleaved Cas 3 and cleaved Parp1) in response to BPA-induced damage was also reduced by Mito-TEMPO treatment in porcine COCs. Interestingly, we confirmed the positive effects of Mela with respect to superoxide production upon BPA exposure during oocyte maturation and also confirmed the reduction in mitochondrial apoptosis in Mela (0.1 µM)-treated porcine COCs. These results provide evidence for the first time that antioxidative effects of Mela on BPA-derived superoxide improve porcine oocyte maturation.</P>