不同浓度锶对MC3T3-E1细胞增殖、ALP活性及成骨分化的影响

Effects of strontium on the proliferation, ALP activity and the differentiation of MC3T3 cells

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 study the effects of strontium concentration on the proliferation, ALP activity and the differentiation of MC3T3-E1 cells. Methods MC3T3-E1 cells were cultured in vitro, and added with strontium of different concentrations. We used MTT to detect the proliferation of MC3T3-E1 celsl, PNPP to detect ALP activity, and RT-PCR to detect m RNA of key proteins of osteoblast differentiation. Results When the concentration of strontium was higher than 500 μg / ml, the proliferation of osteoblasts was significantly decreased, and the growth was inhibited. When the concentration of strontium was 3.91- 250.00 μg / ml, the promoting effect reached the highest level. With the decrease of strontium concentration, the promoting effect was gradually weakened. In the observation range, the effects of strontium on the activity of ALP was obviously dependent. Strontium could significantly increase the ALP activity of MC3T3-E1. When the concentration of strontium was in the range of 3.91- 125 μg / ml, the ALP activity of osteoblasts was significantly increased. The expressions of RUNX2, ALP, Col1 and m RNA gradually increased with the concentration of strontium in a certain range. The expression of m RNA reached the peak value, while the concentration reached 31.25 μg / ml. Conclusions Strontium promotes the proliferation and differentiation of osteoblasts in a dose-dependent manner. However, when the concentration of strontium exceeds the optimal concentration range, its promotion effect is decreased, and the toxicity is inhibited after a certain threshold value.