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目的 观察体外原代培养的人牙乳头细胞内Smad2蛋白的表达 ,以及Smad2蛋白在转化生长因子 β1信号转导中的作用 ,探讨Smad2信号途径在TGF β1调控牙乳头细胞向成牙本质细胞分化过程中的作用机制。方法 原代培养人牙乳头细胞 ,Westernblot杂交方法检测内源性Smad2的表达和TGF β1调控下的表达变化 ,用免疫组化方法观察Smad2细胞内定位变化。结果 培养的人牙乳头细胞表达Smad2蛋白 ,TGF β1能引起Smad2从胞浆转位至核内聚集 ,而BMP 2无此功能 ;TGF β1或BMP 2刺激作用 6h、12h、2 4h和 48h ,牙乳头细胞内Smad2蛋白表达水平未见明显的改变。结论 人牙乳头细胞内存在Smad2基因的表达 ,Smad2能被TGF β1活化后转位至核内发挥作用 ,但Smad2基因的表达无明显变化 ,提示TGF β1在调控牙乳头细胞向成牙本质细胞分化的过程中 ,Smad2信号途径可能发挥一定的作用。
Objective To observe the expression of Smad2 protein in human primary cultured human dental papilla cells and the role of Smad2 in transforming growth factor β1 signal transduction in vitro and to explore the role of Smad2 signaling pathway in regulating the differentiation of human dental papilla cells into odontoblasts In the mechanism of action. Methods Human dental papilla cells were cultured in primary culture. The expression of endogenous Smad2 and the expression of TGFβ1 were detected by Western blot. The localization of Smad2 was observed by immunohistochemistry. Results The cultured human dental papilla cells expressed Smad2 protein. TGFβ1 induced the translocation of Smad2 from the cytoplasm to the nucleus, whereas BMP 2 did not. TGFβ1 or BMP 2 stimulated for 6h, 12h, 24h and 48h No significant changes in Smad2 protein expression in papillary cells. Conclusion The expression of Smad2 gene exists in human dental papilla cells. Smad2 can be activated by TGF β1 and then translocates to the nucleus. However, the expression of Smad2 gene has no obvious change, which indicates that TGF β1 regulates the differentiation of dental papilla cells into odontoblasts Smad2 signaling pathway may play a role.