持续性甲状旁腺素抑制成骨细胞分化的实验研究

Experimental study of the inhibitory effect of continuous treatment with parathyroid hormone in osteoblast differentiation

目的探讨持续性给与甲状旁腺素(PTH)对成骨细胞分化过程的抑制作用。方法培养MC3T3-E1成骨前体细胞,持续性给与PTH处理,碱性磷酸酶(ALP)染色方法,检测细胞内ALP的分泌;免疫荧光方法检测细胞内Osterix(Osx)蛋白的表达;real-time RT-PCT法和Westernblot方法检测细胞内成骨因子基因和蛋白的表达。结果 MC3T3-E1成骨前体细胞具有自发向成骨细胞方向分化的特征,与对照组相比,经PTH持续性处理的细胞ALP染色强度明显降低,Osx免疫荧光强度较弱,细胞内成骨因子基因和蛋白的表达亦显著低于对照组。结论持续性给与PTH具有抑制成骨细胞分化的作用。

Objective To investigate the effect of continuous treatment with parathyroid hormone（PTH） to inhibit the osteoblast differentiation.Methods MC3T3-E1 cells were cultured in the presence of continuous PTH treatment.Alkaline phosphatase（ALP） staining was performed to detect the ALP protein.Osterix（Osx） expression was detected with immunofluorescence.Real-time PCR and Western blotting were performed to examine the gene and protein expressions of osteoblastic factors.Results MC3T3-E1 cells had the characteristic of the spontaneous osteoblast differentiation.Comparing with control group,ALP staining density decreased dramatically in the continuous PTH treatment group.Osx protein expression also decreased.The gene and protein expressions of osteoblastic factors in the treatment group were also lower than those in the control group.Conclusion Continuous PTH treatment inhibits osteoblast differentiation.