重组人骨形态发生蛋白-9对人牙周膜成纤维细胞成骨分化的影响  

Effect of Recombinant Human BMP-9 on Osteogenesis of Human Periodontal Ligament Fibroblasts

作　　者：陆蓓[1] LU Bei(Department of Stomatology,Second People＇s Hospital of Chengdu,Chengdu 610017,China)

机构地区：[1]成都市第二人民医院口腔科,四川成都610017

出　　处：《口腔医学研究》2018年第8期857-861,共5页Journal of Oral Science Research

基　　金：四川省科技厅基金(编号:2015SZ0158)

摘　　要：目的:研究重组人骨形态发生蛋白-9(recombinant human bone morphogenetic protein-9,rhBMP-9)对体外人牙周膜成纤维细胞成骨细胞分化的影响。方法:体外培养人牙周膜成纤维细胞,成骨分化培养基(osteogenic differentiation medium,ODM)诱导成骨分化,加入不同浓度rhBMP-9或p38和胞外信号调节激酶1/2(extracellular signalregulated kinase1/2,ERK1/2)抑制剂SB203580和U0126处理。采用LabAssayTM碱性磷酸酶(alkaline phosphatase,ALP)检测试剂盒检测ALP活性。提取总RNA,实时荧光定量聚合酶链反应(Real time fluorescence quantitative PCR,qPCR)检测RUNT相关转录因子2(Runt-related transcription factor 2,RUNX2)、分化抑制因子1(inhibitor of differentiation factor 1,ID1)、骨唾液酸蛋白(bone sialoprotein,BSP)和骨桥蛋白(osteopontin,OPN)表达情况。结果:rhBMP-9可以显著增加人牙周膜成纤维细胞ALP活性,且呈浓度和时间依赖性。成骨分化相关转录因子RUNX2和ID1以及晚期成骨标志物BSP和OPN的表达在rhBMP-9的刺激下也显著升高。与rhBMP-9单独刺激组比较,加入SB203580和U0126共刺激后可完全抑制rhBMP-9促人牙周膜成纤维细胞成骨分化的作用。结论:rhBMP-9能够显著诱导人牙周膜成纤维细胞向成骨细胞样细胞分化,且其可能机制为调节p38和ERK1/2通路。Objective： To investigate the effect of recombinant human BMP-9 （rhBMP-9） on the differentiation of human periodontal ligament fibroblasts into osteoblasts. Methods： Human periodontal ligament fibroblasts were cultured in vitro and osteogenic differentiation was induced by osteogenic differentiation medium （ODM）. Then the cells were treated with different concentrations of rhBMP-9 in the presence or absence of p38 and extracellular signal-regulated kinase 1/2 （ERK1/2）. Alkaline phosphatase （ALP） activity was measured using LabAssayTM ALP assay kit. The total RNA was extracted and real time fluorescence quantitative （qPCR） was used to detect the expressions of Runt-related transcription factor 2 （RUNX2）,inhibitor of differentiation factor 1 （ID1）,bone sialoprotein （BSP） and osteopontin （OPN）. Results： RhBMP-9 stimulation significantly increased the ALP activity of human periodontal ligament fibroblasts in a concentration and time-dependent manner. Osteogenic differentiation-related transcription factors RUNX2 and ID1, as well as the expression of late osteogenic markers BSP and OPN, were also significantly increased by stimulation with rhBMP-9. Compared with rhBMP-9 alone, the effect of rhBMP-9 on osteogenic differentiation of human periodontal ligament fibroblasts was completely abolished by adding SB203580 and U0126 costimulation. Conclusion： RhBMP-9 could induce the osteogenic differentiation of human periodontal ligament fibroblasts, which might relate to the adjustment of p38 and ERK1/2 pathways.

关 键 词：重组人骨形态发生蛋白-9 成纤维细胞 成骨细胞分化 胞外信号调节激酶1/2 

分 类 号：R781.42[医药卫生—口腔医学]