title: Effect of 7,8-dihydroxyflavone as an antioxidant on in vitro maturation of oocytes and development of parthenogenetic embryos in pigs.
abstract: One of the factors that impairs in vitro produced porcine embryos is the oxidative stress that is mainly caused by the imbalance between reactive oxygen species (ROS) generation and antioxidants activity, especially that of glutathione (GSH). Here, we examined the effect of 7,8-dihydroxyflavone (7,8-DHF), a kind of flavonoid antioxidant, on porcine oocyte maturation and its developmental competence. Porcine oocytes were cultured in media supplemented with 0, 1, 5 and 10 μM 7,8-DHF during both in vitro maturation (IVM) and in vitro culture (IVC) after parthenogenetic activation. Maturation of oocytes was evaluated based on first polar body (PB) extrusion and intracellular GSH level, and developmental competence was assessed through observing cleavage and blastocyst formation. In each step, the levels of intracellular GSH and ROS were assessed by fluorescence intensity, and the apoptosis-related gene expression was examined using semiquantitative RT-PCR. The group treated with 1 μM 7,8-DHF during IVM and IVC showed increased cytoplasmic maturation and reached the blastocysts stage (36.1%) at a higher rate than the other groups (24.7, 16.0 and 10.3% for 0, 5 and 10 μM, P<0.05). In that group, the intracellular GSH level was significantly increased while ROS generation was significantly decreased after IVM and IVC (P<0.05). Moreover, it showed high expression of an anti-apoptotic gene (BCL2L1) and low expression of a pro-apoptotic gene (BAK1) (P<0.05). In conclusion, treatment with 1 μM 7,8-DHF during IVM and IVC showed an anti-apoptotic effect by increasing intracellular GSH synthesis and scavenging ROS and therefore improved the developmental competence of porcine embryos.
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Seoul 151-742, Korea.
authors.forename: Ji-Yei
authors.initials: JY
authors.lastname: Choi
authors.forename: Jung-Taek
authors.initials: JT
authors.lastname: Kang
authors.forename: Sol-Ji
authors.initials: SJ
authors.lastname: Park
authors.forename: Su-Jin
authors.initials: SJ
authors.lastname: Kim
authors.forename: Joon-Ho
authors.initials: JH
authors.lastname: Moon
authors.forename: Islam M
authors.initials: IM
authors.lastname: Saadeldin
authors.forename: Goo
authors.initials: G
authors.lastname: Jang
authors.forename: Byeong-Chun
authors.initials: BC
authors.lastname: Lee
mesh_terms: D000003:Abattoirs; D000818:Animals; D000975:Antioxidants / Q000009:adverse effects / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D051017:Apoptosis Regulatory Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D001755:Blastocyst / Q000187:drug effects* / Q000378:metabolism; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D003593:Cytoplasm / Q000187:drug effects / Q000378:metabolism; D015536:Down-Regulation / Q000187:drug effects; D004477:Ectogenesis / Q000187:drug effects*; D046149:Embryo Culture Techniques / Q000662:veterinary; D005260:Female; D047309:Flavones / Q000009:adverse effects / Q000494:pharmacology*; D005978:Glutathione / Q000819:agonists / Q000378:metabolism; D059471:In Vitro Oocyte Maturation Techniques / Q000662:veterinary; D009865:Oocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects; D010312:Parthenogenesis / Q000187:drug effects*; D017382:Reactive Oxygen Species / Q000037:antagonists & inhibitors / Q000378:metabolism; D034421:Sus scrofa; D015854:Up-Regulation / Q000187:drug effects
chemical_list: C485383:6,7-dihydroxyflavone; D000975:Antioxidants; D051017:Apoptosis Regulatory Proteins; D047309:Flavones; D017382:Reactive Oxygen Species; D005978:Glutathione