1,25(OH)2D3在牙乳头干细胞成骨分化期的作用研究

摘要 目的：研究1,25(OH)2D3在牙乳头干细胞向成骨细胞分化过程中的作用角色。方法：分离培养牙乳头干细胞，培养基中添加不同浓度1,25(OH)2D3，MTT法检测细胞生长速度，流式细胞术检测细胞增殖周期变化，western blot方法检测核因子-k B受体活化剂配体（RANKL）、骨保护素（OPG）和维生素D受体（VDR）的蛋白质表达水平；siRNA技术沉默VDR表达后，检测其对RANKL和OPG蛋白质表达的影响。结果：MTT和流式细胞术检测结果显示1,25(OH)2D3对牙乳头干细胞的增殖没有明显作用；western blot结果显示RANKL、OPG和VDR的蛋白质表达与1,25(OH)2D3浓度具有剂量相关性；siRNA沉默VDR表达后，RANKL和OPG的蛋白质表达水平均有不同程度下降。结论：1,25(OH)2D3通过调节VDR水平影响牙乳头干细胞向成骨细胞方向的分化过程。

A study on functional roles of 1,25 (OH) 2D3 inducing osteogenic differentiation of the dental papilla stem cell

Abstract Objective:To study the functional roles of 1,25 (OH) 2D3 inducing osteogenic differentiation of the dental papilla stem cells.Methods:The dental papilla stem cells were isolated and cultured in medium supplemented with different concentrations of 1,25 (OH) 2D3. MTT assay was used to detect cell growth, and flow cytometry to detect cell cycle. Western blot was used to detect protein expression levels of receptor activator of nuclear factor-k B ligand (RANKL), osteoprotegerin (OPG) and vitamin D receptor (VDR). After siRNA silencing VDR expression, RANKL and OPG protein level were detected. Results:MTT and flow cytometry results showed that 1,25 (OH) 2D3 have no obvious effect on dental papilla stem cell proliferation. Western blot results showed that RANKL, OPG and VDR protein expression level were in a dose correlation with 1,25 (OH) 2D3 concentration. When VDR expression was silenced by siRNA, the protein expression levels of RANKL and OPG were decreased as vary degrees. Conclusion:1,25 (OH) 2D3 affects the osteoblast differentiation process of the dental papilla stem cell by adjusting the VDR expression.