CAJ | 학술논문

目的：研究1,25-二羟基维生素D3[1,25(OH)2D3]在牙乳头干细胞向成骨细胞分化过程中的作用。方法分离培养儿童牙乳头干细胞，培养基中添加1,25(OH)2D3，使其终浓度分别为1、10和100 nmol/L，另设对照组加入等体积生理盐水。采用噻唑蓝（MTT）法绘制细胞生长曲线，流式细胞术检测细胞增殖周期变化，Western blot法检测维生素D受体（VDR）、核因子-κB受体活化剂配体（RANKL）及骨保护素（OPG）蛋白表达水平。采用siRNA技术沉默VDR表达后（siR-VDR组），检测其对RANKL和OPG蛋白表达的影响。结果对照组及1、10、100 nmol/L 1,25(OH)2D3组增殖指数（PIx）分别为49.78±2.03、50.14±1.55、48.81±1.91和48.56±2.15，不同浓度1,25(OH)2D3对牙乳头干细胞的增殖无明显作用（F=3.174，P＞0.05）。对照组牙乳头干细胞中可见一定水平VDR、RANKL和OPG的蛋白质表达；加入1,25(OH)2D3后，VDR、RANKL和OPG蛋白表达水平均有不同程度升高，其中以VDR上升趋势最为显著；转染siR-VDR后，VDR、RANKL和OPG的蛋白质表达水平下降。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。
목적：연구1,25-이간기유생소D3[1,25(OH)2D3]재아유두간세포향성골세포분화과정중적작용。방법분리배양인동아유두간세포，배양기중첨가1,25(OH)2D3，사기종농도분별위1、10화100 nmol/L，령설대조조가입등체적생리염수。채용새서람（MTT）법회제세포생장곡선，류식세포술검측세포증식주기변화，Western blot법검측유생소D수체（VDR）、핵인자-κB수체활화제배체（RANKL）급골보호소（OPG）단백표체수평。채용siRNA기술침묵VDR표체후（siR-VDR조），검측기대RANKL화OPG단백표체적영향。결과대조조급1、10、100 nmol/L 1,25(OH)2D3조증식지수（PIx）분별위49.78±2.03、50.14±1.55、48.81±1.91화48.56±2.15，불동농도1,25(OH)2D3대아유두간세포적증식무명현작용（F=3.174，P＞0.05）。대조조아유두간세포중가견일정수평VDR、RANKL화OPG적단백질표체；가입1,25(OH)2D3후，VDR、RANKL화OPG단백표체수평균유불동정도승고，기중이VDR상승추세최위현저；전염siR-VDR후，VDR、RANKL화OPG적단백질표체수평하강。결론1,25(OH)2D3통과조절VDR수평영향아유두간세포향성골세포방향적분화과정。
Objective To study the functional roles of 1,25(OH)2D3 in osteogenic differentiation of the dental papilla stem cells. Methods The dental papilla stem cells were isolated and cultured in medium supplemented with different con-centrations of 1,25(OH)2D3 (1, 10 and 100 nmol/L). MTT assay was used to detect the cell growth, and flow cytometry was used to detect the cell cycle. Western blot assay was used to detect protein expression levels of receptor activator of nuclear factor-κB ligand (RANKL), osteoprotegerin (OPG) and vitamin D receptor (VDR). After siRNA silencing VDR expression, protein levels of RANKL and OPG were detected. Results MTT and flow cytometry results showed that there were no sig-nificant differences in the cell proliferation between different concentrations of 1,25(OH)2D3 (1, 10 and 100 nmol/L) and con-trol groups (P>0.05). Western blot results showed that there were protein expressions of VDR, RANKL and OPG in control group. The protein expressions of VDR, RANKL and OPG were increased after adding 1,25(OH)2D3, in which the upward trend was the most significant in VDR. After VDR expression was silenced by siRNA, the protein expression levels of VDR, RANKL and OPG were decreased. Conclusion 1,25(OH) 2D3 affects the osteoblast differentiation process of the dental pa-pilla stem cells by adjusting the VDR expression.