3,4-Dihydroxyflavone Acts as an Antioxidant and Antiapoptotic Agent to Support Bovine Embryo Development In Vitro

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概要

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• The effects of two antioxidants, superoxide dismutase (SOD) and the flavonoid 3,4-dihydroxyflavone (DHF), on bovine embryo development in vitro were examined. Blastocyst development, total cell and inner cell mass (ICM) numbers, intracellular levels of reactive oxygen species (ROS), apoptotic indices and gene expression levels were examined before and after treatment of day 2 bovine embryos (≥2-4 cells) with various concentrations of 3,4-DHF or SOD for 6 days. Statistical analysis was performed using analysis of variance, with significance defined at the P<0.05 level. SOD had no significant effect on bovine embryo development at any tested concentration (control, 32.8%; 300 U/ml, 33.9%; 600 U/ml, 24.2%). In contrast, 10 μM 3,4-DHF promoted higher blastocyst development (39.3%) than any other concentration (control, 26.7%; 1 μM, 30.3%; 50 μM, 29.5%; 100 μM, 20.5%). Compared with 300 U/ml SOD, 10 μM 3,4-DHF resulted in significantly higher blastocyst development (44.2%) (control, 31.5%; SOD 300 U/ml, 33.6%). Treatment with 3,4-DHF increased the ICM cell number and reduced intracellular ROS production and apoptotic cell numbers. When O2 tension was decreased from 20% (high tension) to 5% (low tension), embryo development rates were doubled regardless of 3,4-DHF treatment. Under high O2 tension, 10 μM 3,4-DHF treatment may render bovine embryo development similar to a low O2 tension environment. The best blastocyst development was obtained under low O2 tension plus 10 μM 3,4-DHF treatment. The relative expression levels of antioxidant (MnSOD), antiapoptotic (Survivin, Bax inhibitor) and growth-related genes (IFN-τ, Glut-5) were significantly increased after 3,4-DHF treatment, while the expression levels of oxidant (Sox) and apoptotic genes (Caspase-3 and Bax) were reduced. These results suggest that 3,4-DHF may promote the in vitro development of bovine embryos through its antioxidant and antiapoptotic effects.

著者

• KIM Eun

  Mirae Biotech/Jeju National University Stem Cell Research Center

• PARK Se

  Mirae Biotech/Jeju National University Stem Cell Research Center

• Lee Sung

  National Crop Experiment Station

• Yang Byoung

  National Institute Of Animal Science

• Lee Hoon

  Department Of Animal Biotechnology Konkuk University

• Han Young

  Mirae Biotech/jeju National University Stem Cell Research Center

• LEE Keum

  Mirae Biotech/Jeju National University Stem Cell Research Center

• JEON Kilsoo

  Mirae Biotech/Jeju National University Stem Cell Research Center

• CHO Ssang

  Department of Animal Biotechnology, KonKuk University

• KO Mon

  National Institute of Subtropical Agriculture

• Riu Key

  Faculty Of Biotechnology College Of Applied Life Sciences Jeju National University

• Cho Ssang

  Department Of Animal Biotechnology Konkuk University

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