【作者】 张韬文；

【导师】 赵望泓；

【作者基本信息】 兰州大学 ， 口腔临床医学， 2011， 硕士

【摘要】 目的使用全反式视黄酸(all-trans retinoic acid,atRA)诱导腭发育敏感期的小鼠作为模型,利用血清蛋白质组学技术,对血清中差异表达的蛋白质和多肽进行鉴定和验证,探讨腭裂发病机制。方法60只C57BL/6J系孕鼠,妊娠10天给予全反式视黄酸(100 mg/kg),建立小鼠腭裂模型,采集妊娠14天孕鼠血清。实验分为三组：全反式视黄酸组、空白组、植物油对照组。采用双向电泳(2-DE)、偶联基质辅助激光解吸电离—飞行时间质谱(MALDI-TOF MS)技术,寻找腭裂孕鼠血清差异表达蛋白质。并通过Western blot对试验结果进行验证。结果触珠蛋白β-chain(Haptoglobinβ-chain, Hpβ-chain)和血清淀粉样P成分(Serum amyloi d P-component, SAP)在全反式视黄酸诱导腭裂孕鼠血清中表达上调,分别是正常孕鼠血清蛋白含量的2.5和3倍。另外还发现触珠蛋白β-chain在全反式视黄酸组上调表达的同时发生糖基化修饰的变化。Westernblot验证结果与2-DE结果相一致。结论HPβ-chain和SAP在全反式视黄酸致腭裂孕鼠血清中表达上调,为进一步研究全反式视黄酸致小鼠腭裂畸形的病理机制打下基础。同时,发现HPβ-chain在腭裂孕鼠血清中糖基化修饰改变,提示糖基化修饰在全反式视黄酸致腭裂畸形中具有重要调控作用。更多还原

【Abstract】 Objective To explore the protein changes in serum of the cleft palate disease, the animal model of the retinoic acid induced cleft palate and the comparative proteome analysis were used in this study.Methods Sixty C57BL/6J strains of mice were used in this study. For the experiment group, E10 pregnant female mice were given by gastric intubations of a single dose of all-trans retinoic acid (RA) (100 mg/kg) and the RA was dissolved in olive oil. The control group were given the equivalent volume of the carrier alone. The normal group did not give anything. Serum were collected from the pregnant mice at E14.2-DE, MALDI-TOF MS, and Western blotting were applied to analysis the protein changes in the serum of the normal, control, and RA pregnant mice groups.Results Haptoglobin B-chain and serum amyloid P-component levels in sera of RA pregnant mice were significantly higher than those from normal or control subjects.Volum% for Haptoglobin B-chain and serum amyloid P-component were 3 and 2.5 times higher,respectively. In addition, N-glycans of haptoglobin B-chain were found differently altered expression between RA groups and normal or control groups. Each of these differences was statistically significant(p value<0.01),p value obtained by Mann-Whitney U-test.Conclusion The expression level of the protein, haptoglobinβ-chain and serum amyloid P-component, in the serum of RA pregnant mice group were significantly higher than those of the normal and control groups. Differences in structure of the N-glycans of haptoglobin B-chain were found in RA pregnant mice. It was suggested that these potential functional proteins provide insights into the pathogenetic mechanism of cleft palate caused by excess RA remains.更多还原