Screening Differentially Expressed Proteins Of Cleft Palates Induced By 2,3,7,8-Tetrachlorodibenzo-P-Dioxin In Mice

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:⑴Cleft palate mice model was established by exposing pregnant C57BL/6 mice to TCDD. Pregnant mice in experimental group were given TCDD (64μg/kg) with intragastric administration, corn oil was given to pregnant mice with same method in control group on gestational day (GD) 12.GD18, Survival rate of fetus was calculated. Palate of fetus was gain for histology examination.⑵Compare proteomics study on related protein of cleft palate induced by TCDD in mice. Pregnant mice in experimental group were given TCDD (64μg /kg) with intragastric administration, corn oil was given to pregnant mice with same method in control group on (GD) 12. Palate tissues were gain for abstracting total protein on GD18. The total protein was separated by two dimensional gel electrophoresis. The differentially expressed proteins between the two groups were compared using image analysis software. The proteins with significant difference were identified by mass spectrometry. The expression of the differential protein was confirmed by immunohistochemistry.Results:⑴Cleft palate mice model was established successfully by exposing pregnant C57BL/6 mice to TCDD. The incidence of cleft palate in the experimental group was 100%. MES don't contact and fusion, leading to cleft plate in our experimental group. The MES subsequently disappears, leading to a continuous palatal shelf consisting of a mesenchymal core bounded by the nasal and oral epithelium in our control group.⑵The proteins with significant difference were identified by mass spectrometry. They were gamma-actin, Prx 1, GDIs,malate dehydrogenase, unnamed protein product,galectin-7,Triosephosphate isomerase,glyceraldehyde-3- phosphate dehydrogenase,phosphoglycerate kinase,ATP synthase. Prx1 protein was up-regulated in experimental group. Immunohistochemistry showed that, Prx 1 protein of fetal palate was increased in experimental group.Conclusion:⑴Cleft palate mice model was established successfully by exposing pregnant C57BL/6 mice to TCDD. The incidence of cleft palate in the experimental group was 100%. The animal model is suitable to study on aetiology and pathogenesy of cleft palate.⑵Up- regulated Prx 1 pro- tein may be associated with cleft palate in mice induced by 2, 3, 7,8-TCDD: oxidative stress is generated by TCDD, following up-regulated Prx 1 protein, which inhibited MEE apoptosis. Then the development and mutual integration of lateral palatine process were disorder.