[Effects of icariin on the proliferation, differentiation and maturation of rat calvarial osteoblasts in vitro].
OBJECTIVE
To investigate the effects of icariin on the proliferation, differentiation and maturation of rat calvarial osteoblasts (ROB).


METHODS
Segregated neonatal SD rat skull,enzyme digestion to obtain ROB. The culture medium was replaced every three days. Serial subcultivation proceeded when cells covered with 80% culture dish. Icariin was added into the culture at 1 x 10(-4), 1 x 10(-5), 1 x 10(-6), 1 x 10(-7) mol/L respectively. MTT method was adopted in proliferation analysis. The activity of ALP was assayed after 9 days' induced. Optimal concentration icariin was added into the medium, then the osteogenic differentiation markers including mineralized bone nodules, CFU-F(ALP) were compared between the icariin-added group and the control. Total RNA was isolated and the gene expressions of Runx-2 and Osterix were investigated by Real Time RT-PCR. Total protein was also isolated and the secretion of collagen I was examined by Western-blot.


RESULTS
The ROB proliferation was inhibited by icariin in a dose-dependent manner. But it evidently led to osteogenic process and maturation. 1 x 10(-5) mol/L was the best concentration. Icariin improved the secretion of collagen I, CFU-F(ALP) amounts and mineralized nodules significantly. It also enhanced the mRNA level of Runx-2 and Osterix.


CONCLUSIONS
The icariin with final concentration of 1 x 10(-5) mol/L can enhance the osteogenic differentiation and maturation of ROB significantly, suggesting that icariin has the activity of inducing bone formation, it has the potential to be developed into a new drug of anti-osteoporosis.
College of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China. zhaiyk1984@163.com
Yuan-kun
YK
Zhai
Zhi-zhong
ZZ
Li
Ke-ming
KM
Chen
Na
N
Zhang
Guo-zheng
GZ
Cheng
Rui-qing
RQ
Zhu
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D002113:Calcification, Physiologic / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D024042:Collagen Type I / Q000096:biosynthesis / Q000235:genetics; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004365:Drugs, Chinese Herbal / Q000008:administration & dosage / Q000494:pharmacology*; D005419:Flavonoids / Q000008:administration & dosage / Q000494:pharmacology*; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012886:Skull / Q000166:cytology; D013194:Staining and Labeling
D024042:Collagen Type I; D050659:Core Binding Factor Alpha 1 Subunit; D004365:Drugs, Chinese Herbal; D005419:Flavonoids; D012333:RNA, Messenger; D000469:Alkaline Phosphatase; C056599:icariin