Icaritin promotes maturation and mineralization of mouse osteoblast MC3T3-E1 cells through CXCR4/SDF-1 signal pathway

Abstract

<b>Objective:</b> To investigate the effect of icaritin on maturation and mineralization of mouse osteoblast MC3T3-E1 cells and its mechanism. <b>Methods:</b> The cultured MC3T3-E1 cells were divided into blank control group, CXC chemokine receptor type 4 (CXCR4) inhibitor (AMD3100) group, icaritin group, and icaritin plus AMD3100 group. The expression of CXCR4, stromal cell-derived factor 1 (SDF-1) and osteogenesis-related genes and proteins were detected by real-time RT-PCR and Western blotting after drug treatment for 24 h. The alkaline phosphatase (ALP) activity was determined with ALP kit on d3 and d6; calcium nodules were detected by alizarin red staining after drug treatment for 14 d. <b>Results:</b> Real time RT-PCR showed that compared with the blank control group, relative expressions of <i>CXCR4, SDF-1</i> and osteogenesis-related genes in icaritin group were significantly increased (<i>P</i><0.05 or <i>P</i><0.01); After AMD3100 treatment, the relative expression of <i>CXCR4</i> gene was decreased (<i>P</i><0.05). Western blot showed that compared with the blank control group, relative expressions of CXCR4, SDF-1 and osteogenesis-related proteins in the icaritin group were significantly increased (all <i>P</i><0.01), but were decreased after AMD3100 was added (all <i>P</i><0.01). The ALP activity of icaritin group was significantly higher than that of blank control group (all <i>P</i><0.01) on d3 and d6 after drug treatment, while the activity of ALP was significantly decreased after AMD3100 treatment (all <i>P</i><0.01). At d14 after drug treatment, compared with the blank control group, the area of alizarin red staining was increased in the icaritin group, while it was significantly reduced after the addition of AMD3100. <b>Conclusion:</b> Icaritin may promote maturation and mineralization of mouse osteoblast MC3T3-E1 cells through CXCR4/SDF-1 signaling pathway.