甲状旁腺激素对人牙囊前体细胞核因子κB受体激活因子配体和骨保护素mRNA表达的影响

Effects of parathyroid hormone on receptor activator of nuclear factar kappa-κB ligand and osteoprotegerin expression of human dental follicle cells in vitro

目的:研究不同浓度、不同作用时间的甲状旁腺激素(Parathyroid Hormone,PTH)对体外培养的人牙囊前体细胞表达核因子κB受体激活因子配体(receptor activator of nuclear factor kappa-B ligand,RANKL)和骨保护素(osteoprotegerin,OPG)mRNA的影响。方法:原代培养和纯化人牙囊前体细胞,取生长状况良好的第3～5代人牙囊前体细胞,与不同浓度(0.1、1、10、100 ng/mL)重组人PTH共孵育,分别用RT-PCR检测不同浓度PTH作用2 h和100 ng/mL PTH作用不同时间点人牙囊前体细胞RANKL、OPG的mRNA表达,以GAPDH作为内参,进行灰度值比较。结果:随着PTH浓度(0.1、1、10、100 ng/mL)和PTH作用时间(2、4、8 h)的增加,人牙囊前体细胞RANKL mRNA的表达逐渐增高,OPG mRNA的表达逐渐降低。不同浓度组间和不同时间之间相比,RANKL和OPG mRNA表达量均有显著性差异(P<0.05)。结论:PTH可增强体外培养的人牙囊前体细胞RANKL mRNA的表达,降低OPG mRNA的表达。

AIM： To investigate the effects of parathyroid hormone on receptor activator of NF-κB ligand（RANKL）and osteoprotegerin（OPG） expression in human dental follicle cells in vitro.METHODS： Human dental follicle cells were cultured and the third to fifth passage cells were incubated with different concentrations of PTH for different times.Total RNA was then isolated from human dental follicle cells and subjected to RANKL and OPG mRNA assay by reverse transcription polymerase chain reaction（RT-PCR）.The relative expression levels of RANKL and OPG mRNA were normalized to GAPDH gene expression.RESULTS： PTH treatment increased RANKL mRNA expression and decreased OPG mRNA expression in a dose and time dependent manner.CONCLUSION： PTH can increase RANKL expression and inhibiting OPG expression in dental follicle cells.