骨形态发生蛋白7协同蛇床子素促进骨髓间充质干细胞成骨分化的研究

Study on Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells Promoted by Bone Morphogenetic Protein 7 and Osthol

目的观察骨形态发生蛋白7(BMP7)信号通路协同蛇床子素对骨髓间充质干细胞(BMSCs)成骨基因表达的影响。方法取10只1月龄雌性SPF级C57BL/6小鼠,脱颈椎处死后,将骨髓细胞接种在6孔培养板上,置于37℃、5%CO2培养箱中培养。根据不同给药,随机分为蛇床子素组、Noggin组、BMP7组、蛇床子素+Noggin组、蛇床子素+BMP7组和空白对照组。利用实时荧光定量PCR法检测小鼠成骨相关基因的表达量。结果蛇床子素能够促进Bmp2、Bmp4、Bmp6、Bmp7基因表达上调,其中Bmp7的基因表达提高最显著(P<0.05)。骨钙素(OC)、骨涎蛋白(BSP)、成骨细胞特异性转录因子(Osterix)和Runt相关转录因子2(RUNX2)的基因表达在蛇床子素组、BMP7组及蛇床子素+BMP7组中升高,在Noggin组中下降(P<0.05)。蛇床子素可以减弱Noggin蛋白对成骨基因表达的抑制作用,并能上调成骨基因的表达(P<0.05)。结论蛇床子素可以逆转Noggin蛋白对BMP7成骨表达的抑制,并且一定程度上提高BMSCs的成骨表达含量。

Objective To observe the effects of osthol on osteogenic gene expression of bone marrow mesenchymal stem cells(BMSCs) through bone morphogenetic protein 7(BMP7) signaling pathway. Methods 10 female SPF C57 BL/6 mice of 1 month old were killed by neck removal. The marrow cells were inoculated on a six hole culture plate and cultured in a 37℃,5%CO2 incubator. The mice were randomly divided into six groups according to the different treatment: Osthol group, Noggin group, BMP7 group, Osthole+Noggin group, Osthole+BMP7 group, blank control group. The osteogenesis related genes’ expression in six groups were detected by real-time fluorescence quantitative PCR. Results Osthol could significantly promote the up-regulation of gene expression of Bmp2, Bmp4, Bmp6, Bmp7, especially Bmp7(P<0.05). The expression of OC,BSP, Osterix, RUNX2 and other genes increased significantly in Osthol group, BMP7 group and Osthol+BMP7 group, but decreased in Noggin group(P<0.05). Osthol can attenuate the inhibitory effects of Noggin protein on osteogenic gene expression and up-regulate osteoblast gene expression(P<0.05). Conclusions Osthol can significantly promote the osteogenic gene expression of bone mesenchymal stem cells and reverse the inhibitory effect of Noggin protein on the osteogenic gene expression of BMP7.