ABSTRACT
Objective: To study the effects of ursolic acid on the differentiation of cementoblast cell line OCCM-30 cells, and to provide the theoretical basis for the repair of root restoration. Methods: The OCCM-30 cells in logarithmic growth phase were divided into control group and different concentrations of ursolic acid groups. The OCCM-30 cells in ursolic acid groups were treated with different concentrations (0.625, 1.250 and 2. 500 μmol · L-1) of ursolic acid and the cells in control group did not receive any treatment. MTT method was performed to detect the inhibitory rates of proliferation of the OCCM-30 cells in various groups at different time points (24, 48, and 72 h); ALP (alkaline phosphatase) assay was performed to detect the cell differentiation and mineralization; real-time quantitative PCR was used to determine the expression levels of osteopontin (OPN) mRNA during 24 h; the expression levels of OPN protein at 3 and 5 d after administration were detected by Western blotting method. Results: There were no significant differences in the inhibitory rates of proliferation of OCCM-30 cells between control group and different concentrations of ursolic acid groups (P>0. 05). Compared with control group, the ALP activities in the OCCM-30 cells in 2. 500 μmol · L-1 ursolic acid group at 3, 5, and 7 d after administration were significantly increased (P<0. 05). Compared with control group, the expression levels of OPN mRNA and protein in the OCCM-30 cells in different concentrations of ursolic acid groups were significantly increased (P<0. 01). Conclusion: Appropriate concentration of ursolic acid has no effect on the inhibitory rate of proliferation of OCCM-30 cells, but it can promote the differentiation of OCCM-30 cells and up-regulate the expression levels of OPN mRNA and protein.
ABSTRACT
Objective:To investigate the effects of ursolic acid (UA) on the proliferation and differentiation of osteoclasts (OC), and to explore its role in orthodontic force-induced root resorption and its relationship with OC.Methods:The mononuclear / macrophage cells RAW264.7 were induced to the OC.Tacrolimus acid phosphatase (TRAP) staining and bone resorption lacunae observation were used to identify the induction.CCK-8 method was used to select the appropriate concentration of UA for RAW264.7 cell-free biotoxicity and to observe its effect on the proliferation and differentiation of RAW264.7.In experimental groups, UA with gradient concentrations (1.0,2.5,5.0,10.0,20.0and 40.0 μmol·L-1)were added.UA was not added in control group.Results:The TRAP staining and bone resorption lacunae observation showed that after the RAW264.7 cells were induced for 5 d, the TRAP staining positive cells were found;the resorption lacunae were rounded,and oval, etc,the bottom wall was coarser,and the boundary was clear,which indicated that the RAW264.7 cells were successfully differentiated into the osteoclasts.The CCK-8 detection results showed that high concentration of UA (> 10.0 μmol·L-1) significantly inhibited the proliferation of OC;the appropriate concentration of UA (5.0 μmol·L-1) was in the biological safety concentration range and could inhibit the OC proliferation;low concentration of UA (<2.5 μmol·L-1) had no effect.Conclusion:RANKL can induce the differentiation and maturation of RAW264.7 cells.UA is correlated with the proliferation and differentiation of OC;UA has inhibitory effect on OC at the appropriate concentration (5.0 μmol·L-1) in a time-dependent manner.