四氧嘧啶导致小鼠肝脏损伤相关肝脏差异蛋白的研究

Study on Differential Expressions of Liver Tissue Proteins in the Alloxan-Induced Hepatic Injury in Mice

目的:用更为简便的蛋白提取方法提取小鼠肝脏蛋白,寻找四氧嘧啶导致小鼠肝脏损伤差异表达蛋白质,为四氧嘧啶损伤肝脏的机理研究提供理论基础,为保肝药物研究提供理论依据。方法:将小鼠随机分成正常组、模型组,给模型组腹腔注射四氧嘧啶,同时给正常组注射等量生理盐水;小鼠处死后对肝脏进行外观和组织学观察。采用不同浓度丙酮分级沉淀肝脏蛋白,用SDS-聚丙烯酰胺梯度凝胶电泳(SDS-PAGE)实验寻找正常组和模型组的差异表达蛋白质,进一步以基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)检测差异蛋白的肽质量指纹,在蛋白质数据库中检索鉴定差异蛋白的种类。结论:正常组和模型组的水溶性总蛋白和非水溶性总蛋白均有差异表达的蛋白质分子。选取水溶性蛋白50%丙酮沉淀中某一差异蛋白条带进行肽指纹图谱鉴定,检索结果为一未命名蛋白质。

Objective：To research the differential expression of liver tissue proteins in the Alloxan-Induced hepatic injury in mice by using the simpler method to extract the proteins of liver tissue in mice and provide the theoretical study of the mechanism of alloxan-induced hepatic injury.Methods：The mice were divided into two groups.In the model group,the mice were injected with alloxan and in the control group they were injected with 0.9% brine.Then the liver tissues of the killed mice in the two groups were investigated on aspect and histology.Proteins in the liver tissue were deposited into different grades and the attained proteins were separated by SDS-PAGE grads electrophoresis with colloidal staining method.The differential expressions of proteins in the model group and the control group were analyzed and the peptide mass fingerprints were detected with MALDI-TOF-MS.The corresponding proteins were retrieved in Databank.Results：The differential depressions of proteins were found both in the water-soluble proteins and non-water-soluble proteins in the two groups.One of the differential proteins in water-solubility-protein extracted by 50% acetone was choose for further analysis and was found to be an unnamed protein.