先天性白内障小鼠不同发育阶段晶状体蛋白的变化机理及突变体研究

Congenital cataract was a common cause of blindness in children. About 1/3 of congenital cataract is genetic. Genetic factors played a major role in the pathogenesis of congenital cataract. Because of the complexity of cataract disease，it is very difficult to really understand the pathogenesis of this disease and treatment of the disease. There was an important relation in changes of lens protein with cataract formation. The pathological phenomena was caused by change of lens protein or quantity. It was very important for the study of this protein. At present, the lens proteomics have drawn more attention. Proteomics is a new science that studies overall proteinaceous network and its development and will provides the basis in theory and technique for clinical diagnosis and etiopathogenesis. Accordingly, avail oneself of the congenital cataract mice model，lens proteomics of mice in different developmental stages were analyzed by using iTRAQ. In order to To elucidate the the alteration mechanism ，changes in the quantification of lens proteins were determined by mass spectrometry. Differentially expressed proteins were validated by ELISA and Western blotting. It were analyzed that expression and localization of the mutant by using immunohistochemical method , understanding the role of the congenital cataract causing mutants of γS-crystallin. The pathogenesis of cataract was explored at the molecular level. It was to provide an important basis for the prevention and treatment of congenital cataract disease.

先天性白内障是儿童常见的致盲眼病，大约有1/3先天性白内障是遗传性的。遗传因素在先天性白内障的发病机制上起了主要作用。但是由于白内障疾病的复杂性，距离真正了解这一疾病的发病机制和根治该疾病还有很大距离。晶状体蛋白的变化与白内障的形成有着重要联系，白内障的病理改变为晶状体蛋白质或量的改变而引发，因此对该蛋白的研究显得尤为重要。目前，晶状体蛋白质组学的研究更引人关注，其研究成果为疾病临床诊断、发病机制等提供了理论依据和技术基础。据此，利用先天性白内障小鼠模型，使用iTRAQ蛋白定量分析技术，对不同发育阶段晶状体蛋白质组进行分析研究，定量测定晶状体蛋白的变化，阐明变化机理，对发现的差异蛋白进行验证。针对先天性白内障小鼠模型突变产生的突变体，应用免疫组织化学法分析测定突变体的表达和定位，以期了解该突变体的作用。从分子水平探寻白内障的发病机制，为先天性白内障疾病的防治提供重要依据。

白内障是一种多因素引发的眼科疾病，晶状体蛋白是白内障疾病发病的关键因素。利用先天遗传性白内障小鼠模型，进行先天性白内障小鼠模型晶状体蛋白质鉴定和不同发育阶段先天性白内障与正常晶状体的差异蛋白分析，进而探讨先天性白内障疾病形成机制，为白内障疾病发生机制的阐明和相关药物的防治提供重要依据。. 完成了先天性白内障小鼠模型晶状体蛋白的变化机理研究, 进行蛋白质鉴定，测定不同发育阶段晶状体蛋白质和非晶状体蛋白质浓度。比较不同发育阶段正常和白内障小鼠蛋白质浓度变化规律。使用ELISA检测和免疫印迹法(Western Blotting)对发现的差异蛋白进行验证。阐明先天性白内障小鼠模型蛋白质变性及量的变化机理。. 完成了突变体研究,突变体表达及蛋白纯化，以此作为抗原，通过免疫动物后获得特异性的抗体，再以此抗体去探测组织或细胞中的同类的抗原物质。用酶标记抗体与晶状体162个氨基酸的突变体作用，然后加入酶的底物，生成有色的不溶性产物或具有一定电子密度的颗粒，通过光镜，对晶状体内的相应抗原进行定位或定性研究。以期了解该突变体在先天性白内障小鼠模型晶状体蛋白质的作用。