重组人结缔组织生长因子对牙髓细胞增殖与成牙本质分化的影响

Effect of recombinant connective tissue growth factor on proliferation and odontogenic differentiation of dental pulp cells

目的:研究重组人结缔组织生长因子(recombinant connective tissue growth factor,r CTGF)对牙髓细胞(human dental pulp cells,hDPCs)增殖及分化的影响。方法:利用不同浓度(0、1、10、100 ng/mL)rCTGF分别处理牙髓细胞,CCK8法检测牙髓细胞增殖情况;茜素红染色和半定量试验检测细胞矿化结节的形成变化,qRT-PCR测定成牙本质分化相关基因DMP-1、DSPP和OC的表达情况,Western免疫印迹法测定r CTGF刺激牙髓细胞后,ERK1/2信号通路的磷酸化水平。采用SAS 9.3软件包对数据进行统计学分析。结果:高浓度的rCTGF(100 ng/mL)可以促进牙髓细胞增殖;经矿化诱导后,10 ng/mL rCTGF促进牙髓细胞矿化结节形成的效果最好,钙盐沉积量最明显(P<0.05),成牙本质分化相关基因DMP-1、DSPP的表达显著上调(P<0.05)。Western免疫印迹结果显示,10 ng/mL rCTGF刺激牙髓细胞后,p-ERK1/2蛋白的表达升高。结论:rCTGF可能通过激活ERK1/2信号通路,促进牙髓细胞的增殖与分化。

PURPOSE:To analyze the effect of recombinant connective tissue growth factor(rCTGF)on proliferation and differentiation of human dental pulp cells(hDPCs).METHODS:Human dental pulp cells were cultured in vitro and treated with r CTGF at different concentrations(0,1,10,100 ng/mL).The proliferation of dental pulp cells was detected by CCK8 assay.The formation of mineralized nodules was determined by alizarin red staining and half-quantitative alizarin Red S assay.qRT-PCR was utilized to detect the expression of odontogenic differentiation related genes DMP-1,DSPP and OC,and the phosphorylation level of ERK1/2 signaling pathway was detected by Western blot.The data were analyzed with SAS 9.3 software package.RESULTS:High concentration of rCTGF(100 ng/mL)could promote proliferation of dental pulp cells.After mineralization induction,10 g/mL rCTGF had the best effect on promoting the formation of mineralized nodules in dental pulp cells,and calcium ion deposition was the most obvious(P<0.05).The expression of odontogenic differentiation related genes DMP-1 and DSPP was significantly up-regulated(P<0.05).Western blot results showed that hDPCs stimulated by 10 ng/mL rCTGF could increase the expression of p-ERK1/2.CONCLUSIONS:rCTGF may promote the proliferation and differentiation of human dental pulp cells through activating ERK1/2 signaling pathway.