title: Sequential treatment with resveratrol-trolox improves development of porcine embryos derived from parthenogenetic activation and somatic cell nuclear transfer.
abstract: We investigated the effect of resveratrol supplementation during IVM and/or trolox during IVC on the development of porcine embryos derived from parthenogenetic activation (PA) and SCNT. In this study, we evaluated intracellular glutathione (GSH) and reactive oxygen species (ROS) levels, gene expression in blastocysts, and embryonic development after PA and SCNT. To determine the combined effects of resveratrol during IVM and trolox during IVC on PA embryos, we selected optimal concentrations (2 μM of resveratrol and 200 μM of trolox) and designed four groups: (1) control, (2) resveratrol, (3) trolox, and (4) combined. All treatment groups showed significantly increased intracellular GSH levels and decreased ROS levels. Resveratrol supported significantly higher cleavage and blastocyst formation rates than the control (80.3% and 38.0% vs. 71.1% and 22.4%, respectively) by downregulating Bax/Bcl-2, Caspase-3, and Bak. Trolox showed significantly increased blastocyst formation rates (36.7%) compared with the control (22.4%) by downregulating only Caspase-3. The combined group had significantly higher cleavage and blastocyst formation rates and greater total cell numbers than the control (81.7%, 36.3%, and 67.1 vs. 71.1%, 22.4%, and 47.8, respectively) by downregulating Bax/Bcl-2, Caspase-3, and Bak. On the basis of these results, we applied sequential treatments with resveratrol and trolox to SCNT, and blastocyst formation rates and total cell numbers were significantly increased compared with the control (17.2% and 52.1 vs. 11.8% and 36.6, respectively), with increased GSH, decreased ROS levels, upregulated proliferating cell nuclear antigen, and downregulated Bax/Bcl-2 and Caspase-3. These results indicate that sequential treatment with resveratrol during IVM and trolox during IVC improved the development of PA and SCNT porcine embryos by regulating intracellular GSH, ROS levels, and gene expression.
authors.affiliation: Department of Theriogenology and Biotechnology, Research Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University, Seoul, Republic of Korea.
authors.forename: Sanghoon
authors.initials: S
authors.lastname: Lee
authors.affiliation: Department of Theriogenology and Biotechnology, Research Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University, Seoul, Republic of Korea.
authors.forename: Eun Jung
authors.initials: EJ
authors.lastname: Park
authors.affiliation: Department of Theriogenology and Biotechnology, Research Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University, Seoul, Republic of Korea.
authors.forename: Joon Ho
authors.initials: JH
authors.lastname: Moon
authors.affiliation: Department of Theriogenology and Biotechnology, Research Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University, Seoul, Republic of Korea.
authors.forename: Su Jin
authors.initials: SJ
authors.lastname: Kim
authors.affiliation: Department of Theriogenology and Biotechnology, Research Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University, Seoul, Republic of Korea.
authors.forename: Kilyoung
authors.initials: K
authors.lastname: Song
authors.affiliation: Department of Theriogenology and Biotechnology, Research Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University, Seoul, Republic of Korea; Institute of Green Bio Science and Technology, Seoul National University, Pyeongchang, Gangwon-do, Korea. Electronic address: bclee@snu.ac.kr.
authors.forename: Byeong Chun
authors.initials: BC
authors.lastname: Lee
mesh_terms: D000818:Animals; D002839:Chromans / Q000494:pharmacology*; D046149:Embryo Culture Techniques; D004622:Embryo, Mammalian / Q000187:drug effects*; D047108:Embryonic Development / Q000187:drug effects*; D005978:Glutathione / Q000378:metabolism; D053652:Nuclear Transfer Techniques / Q000662:veterinary*; D017382:Reactive Oxygen Species / Q000378:metabolism; D000077185:Resveratrol; D013267:Stilbenes / Q000494:pharmacology*; D013552:Swine / Q000196:embryology*
chemical_list: D002839:Chromans; D017382:Reactive Oxygen Species; D013267:Stilbenes; D005978:Glutathione; D000077185:Resveratrol; C010643:6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid
keywords: Parthenogenetic activation; Porcine embryo; Resveratrol; SCNT; Trolox