3,4-Dihydroxyflavone acts as an antioxidant and antiapoptotic agent to support bovine embryo development in vitro.
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 O(2) tension was decreased from 20% (high tension) to 5% (low tension), embryo development rates were doubled regardless of 3,4-DHF treatment. Under high O(2) tension, 10 µM 3,4-DHF treatment may render bovine embryo development similar to a low O(2) tension environment. The best blastocyst development was obtained under low O(2) 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.
Mirae Biotech/Jeju National University Stem Cell Research Center, Seoul, Korea.
Keum Sil
KS
Lee
Eun Young
EY
Kim
Kilsoo
K
Jeon
Ssang Gu
SG
Cho
Young Joon
YJ
Han
Byoung Chul
BC
Yang
Sung Soo
SS
Lee
Mon Suck
MS
Ko
Key Jung
KJ
Riu
Hoon Taek
HT
Lee
Se Pill
SP
Park
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism; D001755:Blastocyst / Q000166:cytology / Q000187:drug effects* / Q000502:physiology*; D053624:Blastocyst Inner Cell Mass / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D002417:Cattle; D002452:Cell Count; D004477:Ectogenesis / Q000187:drug effects*; D046149:Embryo Culture Techniques; D047309:Flavones / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects; D051296:Glucose Transporter Type 5 / Q000235:genetics / Q000378:metabolism; D007370:Interferon Type I / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D010100:Oxygen / Q000009:adverse effects; D011257:Pregnancy Proteins / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D055747:SOX Transcription Factors / Q000235:genetics / Q000378:metabolism; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism
C559991:3,4'-dihydroxyflavone; D000975:Antioxidants; D051017:Apoptosis Regulatory Proteins; D047309:Flavones; D051296:Glucose Transporter Type 5; D007370:Interferon Type I; D011257:Pregnancy Proteins; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D055747:SOX Transcription Factors; C020947:interferon tau; D013482:Superoxide Dismutase; D010100:Oxygen