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目的观察不同浓度锶(Sr)对MC3T3-E1细胞增殖、碱性磷酸酶(alkaline phosphatase,ALP)活性及成骨分化的影响规律。方法体外培养MC3T3-E1细胞,将浓度不同的Sr作用于成骨细胞系MC3T3-E1细胞,利用MTT法检测细胞增殖活性,PNPP法检测细胞ALP活性,RT-PCR法检测几种成骨标志分子m RNA的表达情况。结果 Sr离子浓度>500μg/ml时,成骨细胞增殖明显降低,生长受到抑制;浓度在3.91~250.00μg/ml时,促进作用达到最高水平;之后随着浓度降低,促进作用逐渐减弱。在观察范围内,Sr对成骨细胞ALP活性的影响呈明显的浓度依赖性,Sr能显著的增加MC3T3-E1的ALP活性,浓度在3.91~125μg/ml范围时,成骨细胞ALP活性显著提高。RUNX2、ALP、Col1等m RNA的表达量随Sr的浓度在一定范围内逐渐升高,Sr浓度为31.25μg/ml左右时,其m RNA表达量达到峰值。结论 Sr促进成骨细胞的增殖和分化具有显著的剂量依赖关系,超过最优浓度范围其促进作用下降,在一定阈值后甚至会出现毒性抑制作用。
Objective To observe the effects of different concentrations of strontium (Sr) on the proliferation, alkaline phosphatase (ALP) activity and osteogenic differentiation of MC3T3-E1 cells. Methods MC3T3-E1 cells were cultured in vitro, and different concentrations of Sr were used to act on osteoblast cell line MC3T3-E1. MTT assay was used to detect cell proliferation activity. ALPP activity was detected by PNPP method. Several osteogenic marker molecules m RNA expression. Results When the concentration of Sr was more than 500μg / ml, the proliferation of osteoblasts was significantly decreased and the growth was inhibited. When the concentration was 3.91 ~ 250.00μg / ml, the promotion effect reached the highest level; then the effect was weakened with the decrease of concentration. In the observation range, the effect of Sr on ALP activity of osteoblasts was significantly concentration-dependent, and Sr could significantly increase the ALP activity of MC3T3-E1. The ALP activity of osteoblasts was significantly increased at the concentration range of 3.91 ~ 125μg / ml . The expression of RUNX2, ALP, Col1 and other m RNAs increased with the concentration of Sr in a certain range. When the concentration of Sr was 31.25μg / ml, the expression of m RNA peaked. Conclusion Sr can promote the proliferation and differentiation of osteoblasts in a dose-dependent manner. The inhibitory effect of Sr on the proliferation and differentiation of osteoblasts decreases more than the optimal concentration, and even after a certain threshold value, it may even cause toxic effects.