Proliferating effects of the flavonoids daidzein and quercetin on cultured chicken primordial germ cells through antioxidant action.
Primordial germ cells (PGCs) are undifferentiated pluripotent stem cells, whose proliferation is influenced by many internal and external factors. In the present study, a PGC-somatic cell co-culture model was established to evaluate effects of the flavonoids daidzein (DAI) and quercetin (QUE) on proliferation of PGCs from embryonic chickens. PGCs were isolated from the germinal ridge of 3.5-4day embryos and cultured in 5% fetal calf serum (FCS)-supplemented Medium 199. PGC subculture was carried out on chicken embryonic fibroblast feeder (CEF) or follicular granulosa cell feeder (GCF) layers. The subcultured PGCs were challenged with flavonoids alone or in combination with a reactive oxygen substance (ROS)-producing system on CEF for 48h. The results showed a better supporting effect of CEF than GCF. Flavonoids (1microg/ml) significantly promoted PGC proliferation, which could be markedly inhibited by ROS. The oxidative damage by ROS was further manifest by decreased superoxide dismutase activity and glutathione levels. In addition, activation of protein kinase A (PKA) by forskolin significantly stimulated PGC proliferation, but PKA inhibitor H89 inhibited the proliferating effects induced by DAI and QUE. These results indicated that cultured PGCs respond to exogenous agents on proliferation and that antioxidant flavonoids could restore the intracellular antioxidant system and promote PGC proliferation via their antioxidant action involving the PKA signaling pathway.
Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, No. 268 Kaixuan Road, Hangzhou, Zhejiang 310029, China.
Xinyan
X
Tang
Caiqiao
C
Zhang
Weidong
W
Zeng
Yuling
Y
Mi
Hongyun
H
Liu
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D002642:Chick Embryo; D018920:Coculture Techniques / Q000379:methods; D017868:Cyclic AMP-Dependent Protein Kinases; D004341:Drug Evaluation; D004958:Estradiol / Q000494:pharmacology; D005854:Germ Cells / Q000166:cytology / Q000187:drug effects*; D005978:Glutathione / Q000378:metabolism; D007529:Isoflavones / Q000494:pharmacology*; D018384:Oxidative Stress / Q000187:drug effects; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011794:Quercetin / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D013234:Stem Cells / Q000187:drug effects; D013482:Superoxide Dismutase / Q000378:metabolism; D013629:Tamoxifen / Q000494:pharmacology; D014969:Xanthine Oxidase / Q000378:metabolism
D000975:Antioxidants; D007529:Isoflavones; D017382:Reactive Oxygen Species; D013629:Tamoxifen; D004958:Estradiol; C004742:daidzein; D011794:Quercetin; D013482:Superoxide Dismutase; D014969:Xanthine Oxidase; D017346:Protein Serine-Threonine Kinases; D017868:Cyclic AMP-Dependent Protein Kinases; D005978:Glutathione