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추출방법에 따른 감귤 과피 유래 Flavonoid의 추출효율 및 항산화 효과에 대한 비교
최찬익 ( Chan Ick Cheigh ),정원근 ( Won Guen Jung ),정은영 ( Eun Young Chung ),고민정 ( Min Jung Ko ),조상우 ( Sang Woo Cho ),이재환 ( Jae Hwan Lee ),장판식 ( Pahn Shick Chang ),박영서 ( Young Seo Park ),백현동 ( Hyun Dong Paik 한국산업식품공학회 2010 산업 식품공학 Vol.14 No.2
The extraction of polyphenol and flavonoid from citrus peel was performed by the ethanol, sugar, hot water (80oC), and subcritical water extraction methods. The maximum yields of total polyphenolic compounds (27.25±1.33mg QE/g DCP, QE and DCP indicate quercetin equivalent and dried citrus peel, respectively) and flavonoids (7.31±0.41mg QE/g DCP) were obtained by subcritical water extraction (SWE) with operating conditions of 190oC, 1300 psi, and 10 min. The yields by SWE were over 7.2-, and 8.5-fold higher than those of total polyphenols (3.79±0.73mg QE/g DCP) and flavonoids (0.86±0.27mg QE/g DCP) obtained using the ethanol extraction, which showed the highest extraction efficiency among tested conventional methods, respectively. Antioxidant activities of extracts obtained by different methods showed no significant differences. However, the relative antioxidant yield per 1 g dried citrus peel by SWE (190oC, 10 min) was over 9.5-fold higher than that by the ethanol extraction.
Quality assessment by using Rhod-2 staining in porcine matured and fertilized oocytes
Ho-Guen Jega,Hyo-Jin Park,Jin-Woo Kim,Seul-Gi Yang,Min-Ji Kim,In-Su Kim,Sang-Min Lee,Bo-You Heo,Joung Jun Park,Deog-Bon Koo 한국수정란이식학회 2018 한국수정란이식학회 학술대회 Vol.2018 No.11
Mitochondrion is an organelle for regulating calcium (Ca2+) homeostasis. Mitochondrial Ca2+ plays important roles on oocyte maturation, fertilization and embryonic development for ATP production. Low quality oocytes have mitochondrial dysfunction, which lead to overloaded Ca2+ in mitochondria. Recently, Rhod-2 is well known as a mitochondrial derived Ca2+ indicator. However, the changes of Rhod-2 in matured or fertilized porcine oocytes have not been reported. Therefore, the aim of study was to identify the effects of mitochondrial Ca2+ using Rhod-2 on quality assessment of matured oocyte and zygotes in pigs. Thus, we classified two groups (group 1: G1, compact COCs and group 2: G2, uncompact COCs) according to differences of cumulus cells amount and cytoplasm morphology in germinal vesicle (GV) stage of porcine COCs. Therefore, we investigated number of Rhod-2 spots in matured and fertilized oocytes from G1 and G2 groups. The Rhod-2 spot numbers were separated into four parts; n<10, 10≤ n < 20, 20 ≤ n < 30, and 30 < n. The Rhod-2 spots number of G2 group had greater than G1 group in part of 20 ≤ n. Additionally, we investigate mean number of Rhod-2 spots from G1 and G2 groups in matured and fertilized oocytes. As a result, we confirmed that average number of Rhod-2 spots in G2 group increased than that of G2 group. Finally, we also measured the Rhod-2 intensity in matured and fertilized oocytes of G1 and G2 groups. Interestingly, the Rhod-2 intensity in G2 group was higher than that of G1 group. (oocyte: p < 0.001 and fertilized oocyte: p < 0.05). These results demonstrated that changes in Rhod-2 spots and intensity were increased in low quality of matured and fertilized oocytes. Therefore, our results suggest that the differences in mitochondrial calcium level are associated with morphological quality of porcine COCs.
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.