| Objective:To study the effects of Polygonatum sibiricum polysaccharide(PSP) on the osteogenic differentiation of mouse bone marrow mesenchymal stem cells and to discuss its mechanism of action.Methods:1. CCK-8 method was used to detect the effect of different concentration of PSP (5mg/L, 10mg/L,25mg/L,50mg/L, 100mg/L) in intervention groups and control group (Omg/L) on BMSCs at 24,48,72,96 hours.2. Alkaline Phosphatase Assay Kit was used to detect the effect of PSP on the activity of alkaline phosphatase in intervention groups and control group after 7 days of Osteogenic induction.3. Staining of alizarin red was used to detect the effect of PSP on the number of mineral node in intervention groups and control group after 21 days of Osteogenic induction.4. Western Blot was used to detect the effect of PSP on the protein expression of Runx2 after 7days of Osteogenic induction.5. Immunohistochemical staining was used to detect the effect of PSP on the protein expression of COL-1 after 7days of Osteogenic induction and the protein expression of OCN after 7days of Osteogenic induction.Results:1. The OD values showed that there were no inhibitory or cytotoxic effects of PSP on BMSCs at doses (5-100mg/L).2.10,25 and 50 mg/L PSP significantly induced the expression of ALP in the BMSCs on day 7, compared with the untreated control.3. More plaques of calcified extracellular matrix could be detected in 25 mg/L PSP on day 28, compared with the untreated control.4. The protein expression of Runx2 was significantly increased by PSP at the concentration of 5-25mg/L, compared with the control group.5. more positive cells of by Immunohistochemical staining of COL-1 and OCN were observed in 25mg/L PSP group.Conclusion:Polygonatum sibiricum polysaccharide stimulate the osteogenic differentiation of BMSCs through increasing the activity of alkaline phosphatase, the protein expression of Runx2, COL-1 and OCN. |
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