(R)-dehydroxyabscisic alcohol β-D-apiofuranosyl-(1ˮ→6’)-β-D-glucopyranoside enhances the osteoblastic differentiation of ST2 cells via the BMP/WNT pathways

Lonicera japonica has been used in traditional Chinese medicine as an important medicinal plant, with the ability to inhibit osteoclast development and bone loss. However, it is not clear which active ingredient exerts these effects. (R)-dehydroxyabscisic alcohol β-D-apiofuranosy l-(1ˮ→6’)-β-D-glucopyranoside (DAG) is an active constituent isolated from Lonicera japonica. In the present study, the ST2 bone marrow stromal cell line was treated by DAG at different concentrations and the osteoblastic differentiation was explored by ELISA assay, Von Kossa staining, Alizarin Red S staining, reverse transcription-quantitative polymerase chain reaction and western blot analysis. The results revealed that DAG promoted osteoblastic differentiation, as evidenced by increasing mineralization and alkaline phosphatase (ALP) activity, as well as the expression of genes encoding bone differentiation markers, including Alp, osteopontin (Opn) and osteocalcin (Ocn). In addition, DAG upregulated the gene expression of bone morphogenetic protein (Bmp)-2, Bmp4, Wnt family member (Wnt)-1, Wnt3 and runt-related transcription factor 2 (Runx2), as well as the protein expression of phosphorylated-mothers against decapentaplegic homolog (Smad) 1, Smad5 Smad8, β-catenin and Runx2 in ST2 cells. The osteogenic effects induced by DAG were attenuated by the BMP antagonist Noggin and the WNT signaling pathway inhibitor Dickkopf related protein-1. The data indicated that DAG promoted the osteoblastic differentiation of ST2 cells, at least partially through regulating the BMP/WNT signaling pathways. This provides scientific rationale for the development of DAG as a treatment for bone loss-associated diseases, such as osteoporosis.