为了探索力生长因子羧基端E结构域的后24个氨基酸组成的短肽(MGF-Ct24E)对成骨细胞生物学活性的影响,通过组织块培养法获得大鼠原代成骨细胞,采用MTT法和流式细胞仪检测细胞的增殖及细胞周期分布情况,基因芯片技术检测细胞基因表达谱,并用定量PCR实验验证芯片检测结果。结果显示MGF-Ct24E组的细胞增殖活性明显高于对照组,且在培养第一天促增殖效果最为显著。细胞周期结果显示MGF-Ct24E显著提高了S期和G2/M期的细胞所占比例。基因芯片检测发现差异表达基因共1397个,其中上调922,下调475,且差异表达的基因主要是关于细胞的增殖分化调节,生长因子结合和活性调节等方面。MGF-Ct24E对成骨细胞的这种增殖分化调控提示MGF-Ct24E在促进骨修复方面有着潜在的应用价值。
To study the effect of the last 24 amino acids peptide(MGF-Ct24E) corredsponding to the C-terminal part of the mechano growth factor(MGF) E domain on bone formation in vitro,primary osteoblasts were obtained from neonatal rat.MTT assay and cell cycle assay were used to study the proliferation and cell cycle stage distribution of osteoblasts.The gene expression profile of osteoblasts after treat with MGF-Ct24E was measured by DNA microarray analysis.Quantitative PCR was used to verify the microarray data.The results suggested that the proliferation activity of MGF-Ct24E group was significantly higher at the first day compared with control.The significantly greater proportion of S and G2/M phase cells appeared in the MGF-Ct24E group than control.Microarray data showed that 1397 genes were identified to be differentially expressed in the MGF-Ct24E group.Among these identified genes,922 genes were up-regulated and 475 genes were down-regulated.Most of the differentially expressed genes were associated with the regulation of proliferation and differentiation,growth factor activity and binding.The effect of MGF-Ct24E on the regulation of cell proliferation and differentiation suggest a potential role in the treatment of bone repair.