Effect of a novel compound from iLycopodium obscurum/iL. on osteogenic activity of osteoblasts in ivitro/i

摘要

We investigated the potential of an extract of Lycopodium obscurum L.;stigmastane-3-oxo- 21-oic acid (SA), to enhance osteogensis of mouse osteoblastic MC3T3-E1 cells. SA at a concentration of 16 μM was found to have no significant effect upon the viability of the cells, thus concentrations of 8 μM and 16 μM of SA were used in all further experiments. Both concentrations of SA had an inhibitory affect upon alkaline phosphatase activity (ALP) after 8 days incubation, however, after 16 days activity was restored to control levels. However Alizarin red S staining showed increased levels of mineralization for both concentrations after 16 days culture. Real time PCR showed inhibition of genes Runx2 and Osterix genes responsible for the up-regulation of ALP. However early time point (8 days) up-regulation of bone matrix mineralization genes OPN and OCN, and late time point (16 days) up-regulation of both Jun-D and Fra-2 mRNA expression was significantly enhanced. These results suggest a potential mechanism of SA in enhancing bone fracture healing is through the up-regulating bone matrix mineralization.