【摘要】 目的探讨外源性抑制Wnt信号通路对骨形态发生蛋白2(BMP-2)诱导的人牙髓细胞(HDPC)向成牙本质分化的影响。方法将HDPC随机分为对照组、单纯诱导组、抑制剂处理组,对照组不处理,单纯诱导组采用BMP-2诱导液处理,抑制剂处理组采用Wnt抑制剂DKK-1及BMP-2诱导液处理。采用蛋白免疫印迹技术检测各组细胞核β-catenin蛋白表达,PNPP偶氮法检测各组细胞碱性磷酸酶(ALP)活性,Von Kossa染色检测钙结节数量,实时荧光定量PCR与蛋白免疫印迹技术检测各组细胞中牙本质基质蛋白(DMP-1)和牙本质涎磷蛋白(DSPP)表达。结果与对照组相比,单纯诱导组、抑制剂处理组中HDPC细胞核β-catenin蛋白表达下调(P均<0. 05),而ALP活性、钙化结节数量、DSPP和DMP-1 mRNA与蛋白表达均上调(P均<0. 05)。与单纯诱导组相比,抑制剂处理组中HDPC细胞核β-catenin蛋白表达下调(P <0. 05),同时ALP活性、钙化结节数量、DSPP和DMP-1 mRNA与蛋白表达上调(P均<0. 05)。结论外源性抑制Wnt信号通路,可促进BMP-2诱导HDPC分化为成牙本质细胞。更多还原

【Abstract】 Objective To investigate the effect of exogenous inhibition of Wnt signaling pathway on the differentiation of human dental pulp cells(HDPCs) into dentin induced by bone morphogenetic protein 2(BMP-2). Methods HDPCs were randomly divided into the control group,induction group,and inhibitor treatment group. The cells in the control group were not treated. The BMP-2 induction solution was used in the induction group,and the Wnt inhibitors DKK-1 and BMP-2 induction solution were used in the inhibitor treatment group. The expression of β-catenin protein in the nuclei of each group was detected by Western blotting. We detected the alkaline phosphatase activity by PNPP azo method. The number of calcium nodules was detected by Von Kossa staining. The expression of dentin sialophosphoprotein(DSPP) and dentin matrix protein(DMP-1) in cells of each group was detected by real-time fluorescence quantitative PCR and Western blotting. Results Compared with the control group,the expression of β-catenin protein in the HDPC nuclei was down-regulated in the induction group and the inhibitor treatment group(P < 0. 05),while ALP activity,the number of calcified nodules,DSPP and DMP-1 mRNA and protein expression were up-regulated(all P < 0. 05). Compared with the induction group,the expression of β-catenin protein in the HDPC nuclei was down-regulated in the inhibitor treatment group(P < 0. 05),while ALP activity,the number of calcified nodules,DSPP and DMP-1 mRNA and protein expression were up-regulated(all P < 0. 05). Conclusion Exogenous inhibition of Wnt signaling pathway can promote the differentiation of HDPCs into dentin induced by BMP-2.更多还原