利用小鼠全胚胎培养研究环磷酰胺致畸作用

Teratogenetic study of cyclophosphamide in mice using whole embryo culture

本文应用小鼠全胚胎培养技术 ,通过妊娠 d8分别 ip5,1 0 ,1 5和 2 0 mg·kg-1环磷酰胺 ( CP) ,研究了该药对器官原基形成期胚胎的致畸作用 ,并对其致畸机理作了初步探索 .给药 4h后取胚胎进行培养 ,于 d 1 0 .5收获胚胎 ,测量其卵黄囊直径 ,头长及颅臀长并记录其大体形态的变化 .结果表明 ,1 5mg· kg-1组尾畸形率最高 ;2 0 mg· kg-1组生长迟缓率最高 .电镜观察所显示的细胞凋亡的形态学变化及 DNA琼脂糖凝胶电泳的结果均提示 CP致畸作用可能与其诱导的细胞凋亡有关 .

Using the mouse whole embryo culture technique, the teratogenesis induced by cyclophosphamide(CP) (5, 10, 15, 20 mg·kg -1 , ip) and its mechanism were investigated in mice on d 8 of gestation. Embryos were explanted and cultured 4 hours after CP administration. On d 10.5, culture was terminated and embryos were examined for yolk sac diameter, head length and crown rump length and teratogenic effects of CP. Significant tail defect was found in CP(15 mg·kg -1 ) group and CP induced growth retardation was most serious in CP 20 mg·kg -1 group. Results from transmission electron microscopy demonstrated typical changes in apoptosis and cell death in CP 15 mg·kg -1 group. Agarose gel electro phoresis showed typical DNA ladders. These findings suggest that CP induced apoptosis and/or cell death be involved in teratogenesis.