FTY720-P promotes the differentiation and maturation of osteoblasts

CHEN Zhi-ying; ZUO Jie-yi; LAN Wan-li; KUANG Yi-zhen; LIU Xiao-chun; HUANG Dong

doi:10.3969/j.issn.1001-9448.2017.13.003

Abstract：Objective To clarify the mechanism of FTY720-P on osteoblast.Methods Primary osteoblasts were obtained from the skull of Wistar neonatal rats.The growth rate and cell cycle were detected by MTT assay and flow cytometry.Alkaline phosphatase assay and calcified nodule staining were used to compare the morphological changes of FTY720-P cells before and after treatment with FTY720-P.The expression of related proteins and genes were detected by ELISA, Western blot and real time RT-PCR.Results MTT showed that FTY720-P had little effect on cell growth.Flow cytometry showed that the cells were arrested at G2 phase and cell cycle was little affected.Compared with the control group, alizarin red staining showed increased and distinct calcified nodules were observed in FTY720-P treatment group.The blue-violet granules were more prominent in control group than the experimental group in the alkaline phosphatase staining.The expression of OCN and IGF was up-regulated, while the expression of ALP was down-regulated in experimental group.The secretion of OCN and IGF were increased in experimental group, and so was the secretion of extracellular collagen type I.Intracellular ALP activity was reduced in experimental group.Conclusion FTY720-P can promote the maturation and mineralization of extracellular matrix of osteoblasts, but has no effect on cell growth and cycle.