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目的:探讨大鼠颅骨缝成骨细胞在张应力作用下Ets1和Cbfa1基因表达的变化,以揭示骨缝牵张成骨的分子机制。方法:分离培养新生大鼠颅缝成骨样细胞,应用四点弯曲细胞加力装置对其施加单一周期的机械张力,并通过SYBR G reen实时定量RT-PCR技术,对Ets1和Cbfa1的基因表达进行检测。结果:Ets1和Cbfa1的mRNA水平分别在机械张应力作用后6 h及12 h内显著增高,随后,mRNA水平逐渐恢复到对照组水平,Ets1先于Cbfa1表达,在加力后表达立即升高,并在0.5h达到最高水平,而Cbfa1则首先经历一个短暂的潜伏期,后表达逐渐升高,在6 h达到最高水平,Cbfa1的最高表达水平约为Ets1的2.58倍。结论:Ets1和Cbfa1在骨缝牵张中对成骨细胞合成分泌骨基质蛋白可能起着不同的调节作用,而它们的功能具有时序性。高频率的牵张(>2次/24 h)更有利于Ets1和Cbfa1的最佳表达。
OBJECTIVE: To investigate the changes of Ets1 and Cbfa1 gene expression in rat calvarial osteoblasts under tensile stress to reveal the molecular mechanism of osteogenesis distraction osteogenesis. Methods: The osteogenic cells of the cranial suture of neonatal rats were isolated and cultured. Four-point bending cells were used to exert single-cycle mechanical tension. The gene expression of Ets1 and Cbfa1 were detected by SYBR G reen real-time RT-PCR Test. Results: The mRNA levels of Ets1 and Cbfa1 were significantly increased at 6 h and 12 h after mechanical stress, respectively. Subsequently, the levels of Ets1 and Cbfa1 were restored to that of the control group. Ets1 was predominantly expressed in Cbfa1, And reached the highest level at 0.5h. However, Cbfa1 first experienced a short incubation period and then gradually increased after 6h, reaching the highest level at 6h. The highest level of Cbfa1 expression was 2.58 times that of Ets1. CONCLUSIONS: Ets1 and Cbfa1 may play different regulatory roles in osteoblast synthesis and secretion of bone matrix proteins during the process of suture stretch, and their function is time-dependent. High-frequency stretch (> 2 times / 24 h) is more conducive to the optimal expression of Ets1 and Cbfa1.