乙醇对小鼠胚胎脏层卵黄囊发育影响的体外研究  

The Development of Mouse Visceral Yolk SacIs Affected by Alcohol Exposurein Vitro

作　　者：许雅君[1] 王秀娟[1] 肖蓉[1] 李勇[1] 

机构地区：[1]北京大学公共卫生学院营养与食品卫生学系,分子毒理与发育分子生物学实验室,北京100083

出　　处：《癌变．畸变．突变》2005年第2期65-70,共6页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：国家自然科学基金(No.30271364);国家重大基础研究发展基金资助项目(No.2001CB510305)

摘　　要：背景与目的 :探讨乙醇对小鼠胚胎脏层卵黄囊发育的影响及其与畸形发生的联系。 材料与方法 :将8.5dCD_1小鼠胚胎于含不同浓度乙醇的培养基中连续培养48h,乙醇染毒浓度分别为0、1.0、2.0和4.0mg/ml。培养结束后 ,于体视显微镜下对脏层卵黄囊和胚胎的形态发育进行评分 ;测定脏层卵黄囊和胚胎总蛋白和DNA含量 ;HE染色检测卵黄囊组织学改变 ;电镜下观察卵黄囊内皮细胞超微形态 ;半定量RT_PCR法检测一系列卵黄囊血管发育相关基因的表达情况。 结果 :乙醇染毒组卵黄囊直径、血管发育、卵黄囊总蛋白和总DNA含量均较对照组降低 ,且呈现剂量_反应关系。卵黄囊发育障碍与胚胎发育迟缓及畸胎发生率存在相关关系。扫描电镜下可见卵黄囊内皮细胞表面微绒毛数量明显减少 ;透射电镜下可见内皮细胞吞噬活性降低 ,出现核固缩 ,发泡 ,线粒体肿胀等凋亡现象。血管发生相关的Flk1和Tie2基因在乙醇染毒组表达下降。 结论 :乙醇可能通过抑制器官发生期脏层卵黄囊的发育诱导胚胎畸形的发生。BACKGROUND&AIM: To explore the effect of alcohol on the development of mouse visceral yolk sac(VYS)during organogenesis period,and the relationship to the teratogenicity of alcohol. MATERIAL AND METHODS: 8.5day CD-1mouse embryos were cultured in vitro for48hours,with different alcohol concentrations of0、1.0、2.0and4.0mg/ml.The developmental status of VYS and embryos were evaluated at the end of the culture.Hematoxylin and eosin(HE)staining was carried out to detect the histological alteration of VYS.Scanning and transmitting electron microscopy were performed to explore the ultrastructural alterations of endoderm cells of VYS.The expressions of a serial of vasculogenesis/angiogenesis related genes were detected with semi-quantitative RT-PCR. RESULTS: A dose-dependent toxicity to the development of VYS was abserved,including reduced yolk sac diameter,blood circulation,and protein and DNA contents.The hypogenesis of VYS agreed with the developmental retardation and malformations of the embryos.Pathological examination revealed that in the ethanol exposure groups,the endoderm layer of VYS was deranged and large intracellular vacuoles were found in the subapical area.Electron microscopy disclosed fewer microvilli and pinocytotic invaginations on the apical surface of endoderm cells.Signs of apoptosis were also found.The expressions of a series of vasculogenesis and angiogenesis related genes were detected with semi-quantitative RT-PCR.Flk1and Tie2gene were repressed by ethanol,which may count for the disturbance of VYS blood vessel formation.Impaired structural and functional development of VYS may contribute to the teratogenic action of ethanol. CONCLUSION: Alcohol exposure in vitro can affect the development of mouse VYS,which may contribute to the teratogenic effect of alcohol.

关 键 词：乙醇 脏层卵黄囊 致畸性 

分 类 号：R114[医药卫生—卫生毒理学]