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目的:研究重组人骨形态发生蛋白(rhBMP-2)和重组人碱性成纤维细胞生长因子(FGF-2)对成骨细胞(MC3T3-E1 Subclone 14)矿化及焦磷酸合成酶(ENPP1)、跨膜蛋白(ANK)和组织非特异性碱性磷酸酶(TNAP)表达的影响,探讨生长因子影响细胞矿化的机制。方法:将MC3T3-E1 Subclone 14细胞分成3组:成骨细胞诱导液(OS)成骨诱导培养组(对照组),OS与rhBMP-2培养组(rhBMP-2组),OS与FGF-2培养组(FGF-2组)。培养12 d后进行ALP活性检测及茜素红染色,实时荧光定量PCR检测矿化相关基因ENPP1、ANK和TNAP表达的差异。结果:rhBMP-2组ALP活性以及钙化结节明显高于对照组,ENPP1、ANK和TNAP均高表达;FGF-2组ALP活性以及钙化结节明显低于对照组,ENPP1和ANK呈高表达,TNAP低表达。结论:rhBMP-2和FGF-2通过调节ENPP1、ANK和TNAP的表达变化来影响骨的矿化。
OBJECTIVE: To investigate the effects of rhBMP-2 and FGF-2 on the mineralization and pyrophosphate synthetase (ENPP1), osteoblast-like cell line (MCF-3) Transmembrane protein (ANK) and tissue non-specific alkaline phosphatase (TNAP) expression of the impact of growth factors affect cell mineralization mechanism. Methods: MC3T3-E1 subclone 14 cells were divided into 3 groups: osteoblast induction liquid (OS) osteogenic induction group (control group), OS and rhBMP-2 culture group Culture group (FGF-2 group). Alkaline phosphatase (ALP) activity assay and alizarin red staining were performed after 12 days of culture. The expression of ENPP1, ANK and TNAP in mineralization-related genes were detected by real-time fluorescence quantitative PCR. Results: The ALP activity and calcified nodules in rhBMP-2 group were significantly higher than those in control group, and ENPP1, ANK and TNAP were both highly expressed. The ALP activity and calcified nodules in FGF-2 group were significantly lower than those in control group, TNAP low expression. Conclusion: rhBMP-2 and FGF-2 affect bone mineralization by regulating the expression of ENPP1, ANK and TNAP.