目的:研究β2肾上腺素能激动剂福莫特罗(Formoterol)对大鼠的体外骨髓间充质干细胞(MMSC)向成骨细胞分化的影响,进而探讨其作用机制。方法:取SD大鼠的骨髓间充质干细胞,用条件培养液诱导分化后分别加入不同浓度药物,在不同时间点采用RT-PCR法检测细胞分化中Runx2和Osterix的mRNA的表达,采用westernblot法检测细胞中MEK和ERK1/2的磷酸化。结果:在细胞分化早期,加入已知浓度10-7mol/L的Formoterol可抑制成骨样细胞细胞特异性转录因子Runx2mRNA表达;在细胞分化晚期,浓度10-7mol/L的Formoterol也可抑制成骨样细胞OsterixmRNA表达。在加入浓度10-7mol/的Formoterol作用30min后,MEK和ERK1/2的蛋白磷酸化表达均下降。结论:β2受体激动剂可抑制MMSC细胞体外向成骨样细胞的分化,并且可抑制MEK和ERK1/2磷酸化的表达。
Objective:To observe the effects of Formoterol(β2-adrenergic receptor-specific agonist)on the differentiation of rat osteoblast-like cells.Methods:Osterix and RUNX2 are osteoblast-specific transcriptional factors that are essential for osteoblast differentiation and bone formation.RT-PCR was used to detect the Osterix and RUNX2 genes expressions during the treatment of different drug dose,the expression of P-MEK and P-ERK1/2 were measured by western blot.Results:①The mRNA expression of cells cultured with a concentration of 10-7mol/L of Formoterol are obviously low.②The protein phosphorylation of ERK and MEK in osteroblasts decreased at 30min treating the cells with Formoterol.Conclusions:In the present study,we examined the molecular mechanism of Osterix and RUNX2 gene expression,and found that Formoterol inhibited Osterix and RUNX2 expression in the mouse osteoblast-like cells,Formoterol-induced decrease of Osterix and RUNX2 expression may occurred in dose-dependent manner through the activation of extracellular signal-regulated kinase pathway.