不同浓度地塞米松对小鼠腭胚突间充质细胞作用规律的研究

Study on the in vitro effects of dexamethasone at different concentrations on the growth of murine embryonic palatal mesenchymal cell

目的研究细胞水平地塞米松(Dex)致腭裂畸形的生物学机制,筛选出影响腭胚突间充质(EPM)细胞生长的地塞米松最适浓度。方法腭胚突间充质细胞进行原代培养,实验组细胞分别以1×10-9、1×10-8、1×10-7和1×10-6 mol.L-1地塞米松加以干预,MTT比色法检测各组细胞的增殖率,碘化丙啶(PI)染色观察各组细胞的凋亡情况。结果在血清质量分数为10%条件下,随着Dex浓度的增加,细胞增殖率降低。第3天达到了地塞米松药物作用的最高峰。Dex浓度为1×10-6 mol.L-1组与对照组相比,各时段的P值均小于0.01。结论地塞米松在加药后第3天药物作用最显著,之后随着药物的代谢,作用减弱;浓度1×10-6 mol.L-1的地塞米松,既有一定程度的抑制EPM细胞生长的作用,又不会导致大量的细胞死亡。其可作为在细胞水平研究地塞米松致畸的生物学机制的最适浓度。

Objective To observe the effect of dexamethasone on the proliferation and apoptosis of embryonic palatal mesenchymal（EPM） cells,and chose a proper concentration of dexamethasone which can effect the ordinary growth of embryonic palatal mesenchymal cells.Methods The primary EPM cells were isolated and cultured in vitro,then we did biological assay.EPM cells were treated with different concentration dexamethasone（1×10-9,1×10-8, 1×10-7 and 1×10-6 mol·L-1） respectively.The proliferation of EPM cells was evaluated using MTT method.Apoptosis was examined quantitatively with fluorescein stain.Results In the condition of blood serum＇s concentration at 10%, optical density step down following the raise of dexamethasone＇s concentration.The effect of dexamethasone got to a summit at 3 days.Inhibition rate of dexamethasone at 1×10-6 mol·L-1 was the highest.Conclusion Dexamethasone at 1×10-6 mol·L-1 can not only inhibit the growth of the EPM cells,but also will not lead to a large number of cells death.Therefore,this concentration can be used as a reference standard in future research.The most significant drug action time of dexamethason appears at the third day after administration,then the effect became weaken following the drug metabolism.