Leonurusine hydrochloride promotes osteogenic differentiation via Wnt/β-catenin pathway and expression of Runx2

Purpose: To investigate the effect of leonurusine hydrochloride (LH) on osteogenic differentiation, and the involvement of Wnt/β-catenin route and runt-associated transcription factor 2 (Runx2) in the process. Methods: Mouse osteogenic precursor cell line MC3T3-E1 was used. The cells were assigned to control and three other groups treated with increasing doses of LH, i.e., 0.1, 1, and 10 μmmol/L. Cellular differentiation was determined in the various groups with respect to mRNA expressions of alkaline phosphatase (ALP), number of calcified nodules, Wnt/β-catenin pathway-related genes, viz, 5 (LRP5),β-catenin, GSK-3β, as well as mRNA expressions of related osteogenic differentiation genes, namely. alkaline phosphatase (ALP), alpha 1 type I collagen (Col1-α), Run×2, and osteogenic-related transcription factor antibody (Osterix) using reverse transcription polymerase chain reaction (RT-PCR). Results: The secretion levels of ALP in cells exposed to LH doses of 1 and 10 μmmol/L were significantly increased, relative to control values (p < 0.05). There were marked increases in mRNA levels of LRP5 and β-catenin in cells treated with LH doses of 1 and 10 μmmol/L compared with the control group, while GSK-3β mRNA expression levels were significantly lower than that in control group(p < 0.05). However, the expression levels of ALP, Col1-α, Run×2, Osterix mRNA in cells exposed to LH at doses of 1 and 10 μmmol/L were significantly higher than the corresponding expression levels in control cells (p < 0.05). Conclusion: Leonurusine hydrochloride may promote osteogenic differentiation and bone mineralization via a mechanism involving activation of the Wnt/β-catenin pathway and up-regulation of Run×2 expression. Keywords: Leonurusine hydrochloride, Wnt/β-catenin, Runx2, Osteogenic differentiation