外源性Hedgehog抑制剂诱导小鼠口下颌-肢体发育不全模型建立的效果观察  

Hedgehog pathway antagonist-induced oromandibular limb hypogenesis in mouse

作　　者：蒋远晶 茅传青 赖永圳 翁云鹏 张师贤[2] 陈伟辉[1] 

机构地区：[1]福建医科大学附属协和医院口腔颌面外科,福州350001 [2]福建医科大学附属口腔医院口腔颌面外科,福州350002

出　　处：《中华口腔医学杂志》2018年第1期36-41,共6页Chinese Journal of Stomatology

基　　金：国家自然科学基金（31070838）

摘　　要：目的 观察Hedgehog抑制剂（GDC-0449）在胚胎发育期间对胎鼠的致畸作用,建立GDC-0449诱导口下颌-肢体发育不全动物模型,为进一步研究其发病机制提供基础.方法 将27只孕鼠按随机数字表法分为1个对照组和8个给药组,每组3只.8个给药组分别在胚胎发育的不同时间点,胚胎8.5 d（embryonic day 8.5,E8.5）、E9.5、E10.5、E11.5、E12.5、E13.5、E14.5及E15.5给予孕鼠单次灌胃150 mg/kg GDC-0449,于E16.5获取孕鼠胚胎,通过体视镜和HE染色观察胎鼠的颅面部和四肢的畸形情况,从中选取与口下颌-肢体发育不全综合征临床表现极为相似的给药组为最佳给药组.确定最佳给药组后重新取6只孕鼠,按随机数字表法分为对照组和最佳给药组,每组3只,于E18.5取孕鼠胚胎,通过阿尔新蓝-茜素红染色和微型CT观察小鼠全身骨骼的发育情况.结果 E8.5、E13.5、E14.5、E15.5给药组胎鼠未见明显畸形,E11.5、E12.5给药组中出现较高比例的腭裂和舌体呈三角形.E9.5、E10.5给药组中发现胎鼠颅面形态发育异常,包括颅顶部发育异常、小下颌、舌体畸形,部分伴有腭裂与第一磨牙缺失,而且还伴发缺指（趾）畸形,与口下颌-肢体发育不全综合征的临床表现极相似.E9.5给药组胎鼠的致死率[48%（20/42）]相对较高,因此选择E10.5给药组构建口下颌-肢体发育不全的动物模型较理想.结论 在E10.5以150 mg/kg GDC-0449灌胃方式诱导胎鼠腭裂、小舌、小下颌及缺指（趾）畸形,是研究口下颌-肢体发育不全综合征的一种较理想的模型.Objective To analysis teratogenic effect of GDC-0449 to fetus and set up the animal model of GDC-0449 induced oromandibular limb hypogenesis in mouse for further research of its pathogenesis. Methods Twenty-seven pregnant Institute of Cancer Research （ICR） mice were randomly divided into：control group,embryonic day 8.5（E8.5）exposed groups,E9.5 exposed groups,E10.5 exposed groups, E11.5 exposed groups, E12.5 exposed groups, E13.5 exposed groups, E14.5 exposed groups and E15.5 exposed groups. Each group had 3 mice. Exposed groups were treated with the Hedgehog pathway antagonist GDC-0449 at a single dose 150 mg/kg by oral gavage from E8.5 to E15.5. At E16.5, embryonic phenotypes were analyzed in detail by stereo microscope and histology. After establish an optimal dysmorphogenic concentration,6 pregnant ICR mice were randomly divided into control group and the optimal group,embryonic phenotypes were analyzed by whole-mount skeletal staining and micro-computed tomography at E18.5.Results The mice were exposed to GDC-0449 on E11.5 and E12.5 had a high incidence of cleft palate. GDC-0449 exposed between E9.5 and E10.5 caused craniofacial and limb dysmorphology, including micrognathia, microglossia, ectrodactylia, partial anodontia and cleft palate. Most interestingly, these are extremely similar to oromandibular limb hypogenesis syndrome. Conclusions The results of this study indicate that GDC-0449 can be used to induce micrognathia,microglossia,ectrodactylia,partial anodontia and cleft palate.This work established a novel mouse model for oromandibular limb hypogenesis.

关 键 词：模型 动物 口颌系统畸形 Hedgehog蛋白质类 口下颌-肢体发育不全 

分 类 号：R-332[医药卫生] R783