未折叠蛋白反应关键基因在小鼠前肢正常发育和异常发生过程中的表达  

Expression of Key Genes of UPR in Normal and Abnormal Limb Development during Mouse Embryogenesis

作　　者：朱勇飞[1] 张天宝[2] 孙鹂[3] 袁红[3] 

机构地区：[1]杭州师范大学医药卫生管理学院,浙江杭州310018 [2]第二军医大学卫生毒理学教研室,上海200433 [3]杭州师范大学医学实验中心,浙江杭州310018

出　　处：《癌变．畸变．突变》2008年第6期437-440,444,共5页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：浙江省教育厅计划项目(20061055);浙江省自然科学基金(Y206360)

摘　　要：背景与目的:研究未折叠蛋白反应(unfolded protein response,UPR)关键基因在小鼠胚胎前肢正常发育和异常发生过程中的表达情况。材料与方法:ICR小鼠受孕后,将其随机分为实验组和对照组,每组各64只。于孕第10d(gestational day10,GD10),经口灌胃1次给予实验组孕鼠80mg/kg的全反式视黄酸、对照组孕鼠给予等体积的大豆油,并分别于GD11～GD18取两组胎鼠的前肢,利用实时荧光定量PCR检测UPR关键基因Atf6、Ire1、Perk、Grp78的表达丰度。结果:除Ire1在GD18未被检出外,UPR中的上述4个主要基因在GD11～GD18正常肢发育过程中均有表达,且在GD13、GD17时间点前后呈现两个表达峰;其中Grp78的表达丰度最高,Ire1的表达丰度最低。而在实验组异常前肢发育过程中GD15之前上述4个基因的表达丰度均高于对照组正常前肢发育过程中的表达丰度(P<0.05);而GD15后均低于对照组正常前肢的表达丰度(P<0.05),呈相对稳定的低水平状态;且在GD12～GD14上述4个基因有一非常明显的表达高峰,其表达时程比对照组正常肢长,且表达丰度远高于对照组正常肢(P<0.05)。结论:全反式视黄酸所致的短肢模型中,UPR关键基因的表达在GD11～GD14显著升高,而在GD16～GD18则受到抑制,推测UPR可能与致畸有关。BACKGROUND AND AIM： To study the expression of the key genes in UPR,which are Atf6, Irel, Perk and Grp78, in normal forelimb and short forelimb formations during mouse embryogenesis. MATERIALS AND METHODS： On gestational day 10 （GD10）, the pregnant mice of the treatment group were fed with 80 mg/kg all-trans retinoic acid,and those of the control group received same volume of soybean oil. The forelimbs of all embryos were harvested during GD11-GD18, and the expression levels of Atf6, Irel, Perk and Grp78 in all samples were measured by real-time quantity reverse transcript polymerase chain reaction （QRT-PCR） . RESULTS： Except Irel that was not expressed on GD18, the key genes of UPR were all expressed in the normal limbs during GD11- GD18, and there were two expression peaks around GD13 and GD17. In the normal limbs, the expression of Grp78 was highest and that of Irel was lowest, and the difference was so obviously. Before GD15, the expressions of Atf6, Irel, Perk and Grp78 in the abnormal forelinrbs were higher than those in the normal limbs, and after GD15, the expression of these four genes in the abnormal forelimbs were lower than those in the normal limbs, and was relatively stable at a low level. In the abnormal limbs, there was a very obvious expression peak of all genes between GD12-GD14; the time course was longer and the expression of this peak was higher than that of the normal limbs. CONCLUSION： During mouse embryagenesis, the expression abundance of key genes of UPR in all-trans retinoie acid-induced short limb malformations was obviously increased during GD11-GD14, and was suppressed during GD16-GD18, so we think UPR may be teratogenie.

关 键 词：未折叠蛋白反应关键基因 胚胎 肢 发育 

分 类 号：R339.35[医药卫生—人体生理学]