The Role Of RANKL In The Regulation Of NFATc1, C-Src And RANK Gene Expression In Osteoclast-like Cells

Background and aims The aim of this study was explored the relationships of different concentration of Receptor Activator of NF-kB Ligand (RANKL) and the gene expression of NFATcl, c-Src and RANK in vitro,in order to find the relationships of the pathogenesis of osteoporosis and gene expression of NFATcl, c-Src and RANK with the concentration of RANKL.Methods We havest bone marrow stromal cell (BMSC) from five weeks old SD rat and cultured in6-well chamber slides. After one day of incubation, the marrow cells were exposed to M-CSF (25ng/ml) and different concentrations of RANKL (0,50,75, and100ng/ml, respectively) for7days. After7days, the expression of NFATcl、c-Src and RANK mRNA was determined by real-time fluorescent quantitative polymerase chain reaction(RT-PCR).Results Compared with M-CSF (25ng/ml)+RANKL (0ng/ml) group, the levels of NFATcl and c-Src mRNA expression were statistically increased in M-CSF (25ng/ml)+RANKL (75,100ng/ml, respectively) groups (p<0.01, and p<0.01, respectively); Compared with M-CSF (25ng/ml)+RANKL (0ng/ml) group, the levels of RANK mRNA expression was statistically increased in M-CSF (25ng/ml)+RANKL (50,75,100ng/ml, respectively) groups (p<0.01,p<0.01and p<0.01, respectively). Compared with M-CSF (25ng/ml)+RANKL (50ng/ml) group, the levels of NFATcl mRNA expression was statistically increased in M-CSF (25ng/ml)+RANKL(75,100ng/ml) group (p=0.013and p<0.01, respectively); Compared with M-CSF (25ng/ml)+RANKL(50ng/ml) group, the levels of c-Src mRNA expression was statistically increased in M-CSF (25ng/ml)+RANKL (75,100ng/ml) groups (p<0.01and p<0.01, respectively); Compared with M-CSF (25ng/ml)+RANKL (75ng/ml) group, the levels of NFATcl, c-Src and RANK mRNA expression was statistically increased in M-CSF (25ng/ml)+RANKL (100ng/ml) group,(p<0.01,p<0.01, and p<0.01, respectively).Conclutions These data suggest that RANKL could regulate the expression of NFATcl, c-Src and RANK mRNA in the marrow culture system. The experiments illustrate NFATcl, c-Src and RANK and other cytokines play an important role in the pathogenesis of osteoporosis.