唑来膦酸对小鼠胚胎成骨细胞增殖和分化的影响

目的研究唑来膦酸对小鼠胚胎成骨细胞体外增殖和成骨分化的影响。方法将小鼠胚胎成骨细胞体外传代培养,使用含不同浓度（1.0、0.1μmol/L）唑来膦酸的成骨诱导培养基干预细胞,不含唑来膦酸的培养基作对照,培养1、3、5 d采用CCK-8试剂盒检测唑来膦酸对成骨细胞增殖的影响;培养14 d行碱性磷酸酶（ALP）染色;培养21 d行茜素红染色;培养7 d,实时定量聚合酶链式反应（Real-time PCR）检测转录因子Runx2、成骨标志物Ⅰ型胶原（Collagen TypeⅠ）、ALP、骨钙素（OCN）基因的表达,免疫印迹法（Western Blotting）检测转录因子Runx2蛋白的表达。结果不同浓度的唑来膦酸干预细胞后,CCK-8实验检测吸光度值随天数增加而升高,各组之间差异无统计学意义（P〉0.05）。随着唑来膦酸浓度的升高,ALP染色和茜素红染色逐渐变浅,Runx2、Collagen TypeⅠ、ALP、OCN基因的表达量降低,Runx2蛋白的表达量降低,差异均有统计学意义（P〈0.05）。结论唑来膦酸在选定浓度（1.0、0.1μmol/L）下不影响成骨细胞的增殖,但对其分化功能的抑制作用随着浓度的增加而增强。

Effects of zoledronate on proliferation and differentiation of MC3T3-E1 cells

Objective To study the effect of zoledronate（ZOL）on the proliferation and differentiation of MC3T3-E1 cells in vitro.Methods MC3T3-E1 cells bought from Shanghai Cell Bank were treated with different doses of ZOL.The final concentrations of ZOL in the experimental groups were 1.0and 0.1μmol/L respectively,and the control group was given the same volume of culture media without ZOL.At day 1,3 and 5 after ZOL intervention,cell counting kit-8 was used to observe the effects of ZOL on proliferation of osteoblasts.The ALP staining was used on the 14 th day,and the alizarin red staining for calcium on the 21 st day.Real-time PCR was used for the detection of Runx2,OB markers collagen typeⅠ,ALP and OCN mRNA expression,and Western blotting was used for the detection of Runx2 protein expression.Results The absorbance values of cell counting kit-8 assay were increased in ZOL groups time-dependently,and those among ZOL groups had no significant difference.With the increases of ZOL concentrations,the ALP staining and the alizarin red staining were decreased.ZOL could down-regulate Runx2,OB markers collagen typeⅠ,ALP and OCN mRNA expression and Runx2 protein expression（P〈0.05）.Conclusion At concentrations of 1.0and 0.1μmol/L,ZOL have no effect on proliferation of MC3T3-E1,but can inhabit the osteogenic activity of MC3T3-E1 cells in a dose-dependent manner.