BMP-7对高糖环境下成骨细胞生物学性能的影响

The effects of BMP-7 on the biological properties of MC3T3-E1 osteoblasts under high glucose concentration in vitro

目的:探讨高糖环境下不同浓度BMP-7对成骨细胞生物学性能的影响。方法:将成骨细胞(MC3T3-E1Subclone 14)在葡萄糖(25 mmol/L)环境下培养,据BMP-7浓度分为0(对照组)、50、100、200 ng/ml(实验组)4组。检测细胞增殖及ALP活性、矿化结节、成骨基因Runx2及OCN mRNA的表达;培养24 h观察细胞骨架的形态变化。结果:BMP-7可显著促进细胞增殖及ALP活性(P<0.05)。矿化结节数量增多、体积增大明显。实验组Runx2、OCN mRNA水平明显上调(P<0.05);200 ng/ml BMP-7组Runx2、OCN mRNA的水平较其他组同一时间点显著上调(P<0.05)。细胞骨架渐呈网格状铺展且随BMP-7浓度增高而明显。结论:BMP-7可抑制高糖对MC3T3-E1的作用,提示BMP-7有利于促进糖尿病患者种植体的骨结合。

Objective： To investigate the effects of bone morphogenetic protein 7 （BMP-7） on the biological properties of osteoblasts exposed to high glucose. Methods： MC3T3 cells were cultured in the high glucose condition and were divided into 4 groups according to BMP-7 concentration （ng/ml） ： 0 （control group） ,50,100 and 200 ng/mL BMP-7 （test groups）. The cell proliferation and ALP activity were examind 1,3,5 and 7 d after exposure. The number of intracellular calcium nodules were observed with Alizarin red staining after osteogenesis induction for 21 days. The F-actin cytoskeleton of MC3T3 was stained with Rhodamine and examined 24 h after culture. The mRNA expression of OCN and Runx2 were quantified by RT-qPCR 48h after exposure to BMP-7. Results： BMP-7 significandy promoted proliferation of MC3T3 ceils and enhanced ALP activity（P 〈 0.05 ）, increased the number and volume of intracellular calcium nodules. In the cells exposed to BMP-7, the osteoblast form was improved and F-actin cytoskeieton started to change with net- work structure. Compared with control group, the mRNA levels of OCN and Runx2 were up-regulated in BMP-7 groups（P 〈0.05 ）. The mRNA levels of OCN and Runx2 were the highest in 200 ng/ml BMP-7 group （ P 〈 0.05 ）. Conclusion： BMP 7 may inhibit the action of the high concentration of glucose on MC3T3 cells. BMP-7 may be benificial to osseointegration of dental implant in patietents with diabetics.