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

作　　者：曹颖 彭伟伟[1] 王璇 伍甜甜[1] 杜嵘[1] 朱亚琴[1] CAO Ying;PENG Wei-wei;WANG Xuan;WU Tian-tian;DU Rong;ZHU Ya-qin(Department of General Dentistry,Shanghai Ninth People's Hospital,College of Stomatology,Shanghai Jiao Tong University School of Medicine,National Clinical Research Center for Oral Diseases,Shanghai Key Laboratory of Stomatology&Shanghai Research Institute of Stomatology.Shanghai 200011,China)

机构地区：[1]上海交通大学医学院附属第九人民医院·口腔医学院口腔综合科国家口腔疾病临床医学研究中心上海市口腔医学重点实验室上海市口腔医学研究所

出　　处：《上海口腔医学》2020年第1期7-12,共6页Shanghai Journal of Stomatology

基　　金：国家自然科学基金（81271134、81700949）

摘　　要：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.

关 键 词：重组人结缔组织生长因子 牙髓细胞 细胞增殖 细胞分化 ERK1/2信号通路 

分 类 号：R781.3[医药卫生—口腔医学]