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目的 :研究 1,2 5 (OH) 2 D3 对培养人牙乳头细胞Calbindin -D2 8K(CaB)表达和矿化能力的影响。方法 :用 10 -8mol/L 1,2 5 (OH) 2 D3 矿化液处理长期培养的第 2 8天细胞 2 4h ,用放射免疫和免疫组化方法 ,观察CaB表达和碱性磷酸酶 (ALP)、骨钙蛋白 (OC)的变化。同位素示踪方法观察 1,2 5 (OH) 2 D3 对胞外基质4 5Ca2 + 的影响。结果 :加入1,2 5 (OH) 2 D3 后ALP活性约为对照组的 4倍 ;OC含量约为对照组的 1.5倍。CaB在培养 2 8d牙乳头细胞及其胞外基质中表达 ,而在培养 14d牙乳头细胞中CaB免疫反应阴性。 1,2 5 (OH) 2 D3 可加强CaB表达 ,并使胞外基质对4 5Ca2 + 摄取量增加 1.5倍。结论 :1,2 5 (OH) 2 D3 有促进人牙乳头细胞矿化作用 ,表现在不仅能刺激OC、ALP的活性 ,而且可刺激具有分化表型的牙乳头细胞中CaB的合成 ,增加基质对Ca2 + 的摄取。证实促进牙乳头细胞的钙转导是CaB在矿化过程中的作用之一 ,推测CaB被“俘获”在基质中时有诱导Ca2 + 聚集和直接贮存的作用。
Objective: To investigate the effect of 1,25 (OH) 2 D3 on Calbindin-D2 8K (CaB) expression and mineralization in cultured human dental papilla cells. Methods: The long-term cultured cells were treated with 10 -8 mol / L 1,2 5 (OH) 2 D3 mineral solution for 24 hours. Radioimmunoassay and immunohistochemistry were used to observe the expression of CaB and the activity of alkaline phosphatase ALP), osteocalcin (OC) changes. The effect of 1,25 (OH) 2 D3 on 45Ca2 + extracellular matrix was observed by isotope tracer method. Results: After adding 1,25 (OH) 2 D3, the ALP activity was about 4 times that of the control group. The content of OC was about 1.5 times of the control group. CaB was expressed in 28 d dental papilla cells and extracellular matrix, while CaB immunoreactivity was negative in 14 d dental papilla cells. 1,25 (OH) 2 D3 enhanced CaB expression and increased 1.5x extracellular matrix uptake of 4 5Ca2 +. CONCLUSION: 1,25 (OH) 2 D3 can promote the mineralization of human dental papilla cells, which not only stimulates the activity of OC and ALP, but also stimulates the synthesis of CaB in dental papilla cells with differentiated phenotype, Ca2 + uptake. Proved that promoting calcium transduction of dental papilla cells is one of the roles of CaB in mineralization. It is speculated that CaB can induce Ca2 + accumulation and direct storage when it is “captured” in the matrix.