摘要
目的 通过建立小鼠腭裂模型观察全反式维甲酸（all-trans retinoic acid,atRA）对胎鼠腭突间充质细胞增殖及转化生长因子β（transforming growth factor-β,TGF-β）/Smad信号通路的影响,探讨atRA诱发腭裂的机制.方法 将孕鼠根据体质量按随机数字表随机分为atRA组和对照组（每组各27只）,于妊娠第10天（gestation day 10,GD10）上午8时,分别予以100 mg/kg atRA和等体积玉米油溶剂灌胃.HE染色观察GD13-GD15胎鼠上腭发育,5-溴脱氧尿嘧啶核苷（5-bromo-2-deoxyuridine,BrdU）免疫组织化学染色观察atRA对GD13-GD15胎鼠腭突间充质细胞增殖的影响,蛋白质印迹法检测胎鼠腭突间充质细胞内Smad2、磷酸化Smad2（phospho-Smad2,p-Smad2）、Smad4、Smad7的蛋白表达.结果 与对照组相同时间点相比,atRA组GD13-GD15胎鼠腭突间充质细胞BrdU阳性率均显著降低（P〈0.05）,尤其是GD14,atRA抑制率达（65.4±1.7）%.与对照组相同时间点相比,atRA组GD14和GD15胎鼠腭突间充质细胞p-Smad2和Smad4蛋白表达显著降低（P〈0.05）,GD13-GD15胎鼠腭突间充质细胞Smad7蛋白表达显著升高（P〈0.05）.结论 atRA可能通过抑制Smad信号通路的激活影响腭突间充质细胞增殖,从而导致腭裂的发生.
Objective To investigate the mechanism of cleft palate in mice induced by excessive all-trans retinoic acid （atRA）. Methods The pregnant mice were randomly divided into atRA-treated group （n=27） and control group （n=27）. atRA-treated group was given by gavage a single dose of atRA （100 mg/kg） at 8：00 AM on gestation day 10 （GD10） and the control group was given by gavage the isopyknic corn oil. At GD13-GD15, the fetal mice palate development was observed by HE staining. The mouse embryonic palatal mesenchymal cell proliferation was detected by 5-bromo-2-deoxyuridine （BrdU） immunohistochemistry. The expressions of Smad2, phospho-Smad2 （p-Smad2）, Smad4 and Smad7 in mouse embryonic palatal mesenchyme were analyzed by Western blotting. Results At GD13-GD15, compared with the control, the ratio of BrdU-positive cells in the palatal mesenchyme of atRA-treated fetuses decreased significantly （P〈0.05）, especially at GD14, atRA inhibition rate was （65.4 ± 1.7）%. Moreover, atRA decreased the levels of p-Smad2 and Smad4 in embryonic palate mesenchymal cells, whereas the expression of Smad7 was significantly increased at GD14 and GD15. Conclusions atRA may lead to cleft palate by inhibiting the activation of Smad signaling pathway and affecting the proliferation of palatal mesenchymal cells.
出处
《中华口腔医学杂志》
CAS
CSCD
北大核心
2017年第11期690-694,共5页
Chinese Journal of Stomatology
基金
国家自然科学基金（U1304327、21577119、U1604183）
