2,3,7,8-四氯二苯二噁英诱导腭裂发生的差异蛋白筛选

Screening differentially expressed proteins of cleft palates induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin in mice

目的:以蛋白质组学方法筛选2,3,7,8-四氯二苯二噁英(2,3,7,8-tetrachlorodibenzo-p-dioxin,TCDD)致先天性腭裂发生的差异表达蛋白。方法:以TCDD诱导建立胎鼠先天性腭裂模型,行腭部的大体解剖及组织学检查。取实验组和对照组胎鼠的腭组织,分别提取组织总蛋白。通过双向电泳和质谱分析,筛选出实验组和对照组腭组织的差异表达蛋白,并用免疫组化对鉴定出的相关蛋白质进行验证。结果:成功诱导胎鼠腭裂模型。筛选出10种差异表达的蛋白,其中7个蛋白点在试验组上调,3个蛋白点在实验组下调。过氧化物还原酶1(Peroxiredoxin 1,Prx 1)与鸟嘌呤核苷酸解离抑制剂(GDP-dissociation inhibitor,GDI)在实验组表达明显增强。结论:Prx 1、GDI表达的明显增强可能与TCDD所致胎鼠先天性腭裂有关。

Objective：To identify the differentially expressed proteins of 2,3,7,8-tetrachloro-dibenzo-p-dioxin（2,3,7,8-TCDD）induced cleft palate by using two-dimensional gel electrophoresis（2-DE） and mass spectrometry.Methods：On E12,2,3,7,8-TCDD（64 μg/kg） and corn oil（control group） were administered to time-pregnant C57BL/6J mice.Anatomical and histological changes of palates in mouse fetuses were studied on E18.Total proteins were extracted from palate tissue of fetuses from both groups.The total proteins were separated by two-dimensional gel electrophoresis.The differentially expressed proteins between the two groups were compared using image analysis software.The peptide mass fingerprintings（PMF） was acquired after matrix assisted laser desorption/ionization time of flight mass spectrometry（MALDI-TOF-MS）and the proteins were identified by data searching in the Mascot database.The expression of the differential protein was confirmed by immunohistochemistry.Results：Incidence of cleft palate in the experimental group was 100%.Comparative analysis of 2-DE maps revealed 34 differentially expressed proteins between the two groups.Ten differential proteins were further identified by mass spectrometry,of which 7 showed significantly higher volumes in experimental group than in control group and 3 showed significantly higher volumes in control group than in experimental group.Peroxiredoxin 1 protein and guanine nucleotide dissociation inhibitors were up-regulated in experimental group.Conclusion：Up-regulated peroxiredoxin 1 protein and guanine nucleotide dissociation inhibitors may be associated with cleft palate in mice induced by 2,3,7,8-TCDD.