BMP4对人根尖牙乳头干细胞增殖、迁移及矿化的作用机制研究  

Mechanism of BMP4 on the proliferation,migration and mineralization of human apical papilla stem cells

作　　者：贾靖 朱惠芳 张洁 JIA Jing;ZHU Hui-fang;ZHANG Jie(Department 1 of Orthodontics,The Third People's Hospital of Puyang,Puyang 457000,China)

机构地区：[1]濮阳市第三人民医院正畸一科,濮阳457000

出　　处：《口腔颌面修复学杂志》2023年第5期329-334,341,共7页Chinese Journal of Prosthodontics

摘　　要：目的:观察骨形态发生蛋白4(bone morphogenetic proteins 4,BMP4)对人根尖牙乳头干细胞(stem cells from the apical papilla,SCAPs)增殖、迁移及矿化的影响,并探讨可能机制。方法:取P3代对数期SCAPs,随机分为空白组、NC组、mimics组、mimics+DAPT[神经源性基因Notch同源蛋白1(neurogenic locus notch homolog protein 1,Notch1)/Jagged1信号通路抑制剂]组。空白组不处理,NC组转染阴性对照质粒BMP4 NC,mimics组转染过表达质粒BMP4 mimics,mimics+DAPT组转染BMP4 mimics并加入DAPT(10μmol/L)。MTT法检测细胞增殖能力;Transwell实验检测细胞迁移能力;茜素红染色法检测细胞矿化能力;实时荧光定量聚合酶链反应(real-time quantitative polymerase chain reaction,qRT-PCR)法检测细胞碱性磷酸酶(alkaline phosphatase,ALP)、骨涎蛋白(bone sialoprotein,BSP)、骨钙素(osteocalcin,OCN)mRNA表达量;Western blot法检测细胞Notch1、Jagged1、发状分裂相关增强子1(Hes1)蛋白表达量。结果:与空白组、NC组比较,mimics组细胞24 h、48 h、72 h吸光度值,迁移量,矿化结节面积比,ALP、BSP、OCN mRNA表达量,Notch1、Jagged1、Hes1蛋白表达量升高(P<0.05);与mimics组比较,mimics+DAPT组细胞24 h、48 h、72 h吸光度值,迁移量,矿化结节面积比,ALP、BSP、OCN mRNA表达量,Notch1、Jagged1、Hes1蛋白表达量降低(P<0.05)。结论:过表达BMP4可提升SCAPs增殖、迁移能力并促进其矿化,推测其作用机制可能与激活Notch1/Jagged1信号通路有关。Objective:To observe the effect of bone morphogenetic protein 4(BMP4)on the proliferation,migration and mineralization of human apical papilla stem cells(SCAPs),and to explore the possible mechanism.Methods:Duiring the log-phase,SCAPs of P3 generation were randomly divided into blank group,NC group,mimics group,mimics+DAPT[neurogenic gene Notch homologous protein 1(Notch1)/Jagged1 signaling pathway inhibitor]group.The blank group was not treated,the NC group was transfected with the negative control plasmid BMP4 NC,the mimics group was transfected with the overexpression plasmid BMP4 mimics,and the mimics+DAPT group was transfected with BMP4 mimics with the addition of DAPT(10μmol/L).Cell proliferation was detected by MTT assay.Cell migration ability was analyzed by transwell assay.Alizarin red staining was used to test the mineralization capacity of cells.The mRNA expressions of alkaline phosphatase(ALP),bone sialoprotein(BSP)and osteocalcin(OCN)were detected by real-time quantitative polymerase chain reaction(qRT-PCR).Western blot was conducted to analyze the protein expressions of Notch1,Jagged1 and hairy division-related enhancer 1(Hes1).Results:Compared with the blank group and NC group,the absorbance value at 24 h,48 h,and 72 h,migration amount,mineralized nodule area ratio,ALP,BSP,OCN mRNA expression,Notch1,Jagged1,Hes1 protein expression were increased in mimics group(P<0.05).Compared with the mimics group,the absorbance value at 24 h,48 h,and 72 h,migration amount,mineralized nodule area ratio,ALP,BSP,OCN mRNA expression,Notch1,Jagged1,and Hes1 protein expression were decreased in mimics+DAPT group(P<0.05).Conclusion:Overexpression of BMP4 can enhance the proliferation,migration and mineralization of SCAPs.It is speculated that the mechanism may be related to the activation of Notch1/Jagged1 signaling pathway.

关 键 词：骨形态发生蛋白4 根尖牙乳头干细胞 增殖 迁移 矿化 神经源性基因Notch同源蛋白 

分 类 号：K780.2[历史地理—历史学]