[Study on mechanism of the anti hypoxia effect of genistein on osteoblasts in vitro].
OBJECTIVE
To study the protective effect of genistein on osteoblasts treated with hypoxia.


METHODS
Rat osteoblasts were isolated from calvarias of newborn Sprague-Dawly rat by enzyme digestion and hypoxic environment was made by triple-gases incubator. Rat osteoblasts treated with hypoxia for 36 haurs. After 36 hours, cell viability, content of reactive oxygen species (ROS), analysis of cellular cycle and apoptosis, expression of proliferating cell nuclear antigen (PCNA), activity of iNOS and area of calcified nodules were detected. Total RNA was isolated and the gene expression of hypoxia inducible factor-1alpha (HIF-1alpha), BCL-2 and Caspase-3 was investigated by Real Time RT-PCR.


RESULTS
Genistein could significantly improve cell viability, percentage of G1 phases, area of calcified nodules and decrease apoptosis rate, ROS content, expression of PCNA, activity of iNOS. Besides, mRNA levels of HIF-1alpha and BCL-2 were enhanced and that of Caspase-3 was inhibited.


CONCLUSIONS
Genistein can protect osteoblasts from hypoxia and enhance osteogenic differentiation significantly.
Institute of Orthopedics, Lanzhou General Hospital, Lanzhou Command, PLA. Lanzhou 730050, China. jnhanqiu@163.com
Gui-Qiu
GQ
Han
Bao-Feng
BF
Ge
Ke-Ming
KM
Chen
Hui-Ping
HP
Ma
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002453:Cell Cycle / Q000187:drug effects; D015687:Cell Hypoxia; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D005260:Female; D019833:Genistein / Q000494:pharmacology*; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics / Q000378:metabolism; D008297:Male; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism; D012886:Skull / Q000166:cytology*
D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D018809:Proliferating Cell Nuclear Antigen; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D019833:Genistein