叶酸对小鼠腭突间充质细胞增殖的影响

Effects of folic acid on the proliferation of embryonic palatal mesenchymal cells of mouse

目的:从细胞水平对维A酸致小鼠腭裂畸形作用和叶酸是否有拮抗维A酸的致畸作用及其机制进行研究。方法:给予孕鼠过量维A酸,诱导胚胎腭裂模型,不完全消化法提取孕期第14天、17天(GD14、17)的胚胎腭突间充质细胞(EPMcells)进行培养并鉴定细胞;采用四甲基偶氮唑盐(MTT)法检测加入不同浓度叶酸与未加入叶酸细胞增殖的情况。采用SPSS16.0软件包对数据进行统计学处理。结果:①在GD10、GD12管饲孕鼠50mg/kg维A酸,可致胎鼠腭裂畸形;②不完全消化法提纯EPM细胞,纯度达到98%;③维A酸导致GD14的EPM细胞增殖受到抑制,20μg/mL、40μg/mL浓度叶酸可拮抗这种抑制作用。结论:①过量维A酸可致小鼠腭裂畸形,腭突间充质细胞增殖受抑制是其致畸机制之一;②叶酸可拮抗维A酸对小鼠GD14EPM细胞增殖的抑制作用。

PURPOSE： To study the mechanism by which retinoic acid （RA） can cause mouse cleft palate and folic acid could antagonize the effect of RA. METHODS： Pregnant mice were given excess dose of RA to induce cleft palate, and then their embryonic palatal mesenchymal （EPM） cells were extracted by the method of incomplete digestion and cultured at gestation days 14, 17 （GD14, 17）. After being identified, the cells were treated with different concentration of folic acid and then their proliferation was tested by MTT method. SPSS 16.0 software package was used for data analysis. RESULTS： ① Giving pregnant mice 50 mg/kg RA at GD10, 12 could cause mice cleft palate; ② The purity of EPM cells treated by incompleted digestion reached 98%; ③The proliferation of EPM cells of GD14 was inhibited by RA, but folic acid at the concentration of 20 μg/mL, 40 μg/mL could antagonize the effect of RA. CONCLUSIONS： ①Excess dose of RA could cause cleft palate in mouse and an important mechanism is the inhibition of EPM cells’ proliferation; ②Folic acid could antagonize the inhibition effect caused by RA. Supported by Natural Science Foundation of Department of Science and Technology of Shanxi Province （201311056-5）.