| [摘 要] 目的 以双向凝胶电泳和基质辅助激光解析电离飞行时间质谱分析技术为手段，研究透明晶状体与核性先天性白内障混浊晶状体组织差异蛋白质，为核性先天性白内障的致病机制研究提供线索和依据。 方法 从临床手术中抽取核性先天性白内障及与之年龄匹配的透明晶状体的组织样本，抽提晶状体组织总蛋白，采用双向凝胶电泳技术分离蛋白质组分，分析二维凝胶图谱，对比透明晶状体与核性先天性白内障的双向凝胶电泳图谱差异。将稳定出现的差异蛋白斑点原位酶解，经过基质辅助激光解析电离飞行时间质谱仪的肽质量指纹谱测定及相关蛋白质鉴定数据库检索，发现差异蛋白，探寻与核性先天性白内障发病相关的特异蛋白质。 结果 提出了一套从临床手术人晶状体组织蛋白质取材到差异蛋白质分析的方法。分析透明晶状体与核性先天性白内障的双向凝胶电泳图谱，在晶状体蛋白分子量20-30KDa ,等电点4-5的范围内，发现二者晶状体组织之间存在1个差异表达的蛋白,经质谱鉴定及数据库检索确定为晶状体蛋白βA1/3。 结论 通过蛋白质组学研究技术，确定透明晶状体组织与核性先天性白内障混浊晶状体组织中晶状体蛋白βA1/3存在差异表达，该发现为核性先天性白内障的致病机制研究提供线索和思路，对先天性白内障的基础理论进展具有一定意义。
[关键词] 蛋白质组学;核性先天性白内障;双向电泳；质谱；
Proteomic comparison between human congenital nuclear cataract and transparent lens
YU Mei ，ZHU Si-quan， Beijing Tongren Ophthalmic Center,Capital University of Medical Sciences.Beijing 100730,China
Corresponding autther: ZHU Si-quan,Email:zhsq@trhos.com
[ Abstract ] Objective The aim of the study was to comparatively analyze crystallin fragments in congenital cataracts (with nuclear opacity) and age matched transparent lenses to determine the identity of crystallin species that show congenital cataract specific
changes. Methods the protein of transparent lens tissues and opacity lens tissue were separated by 2DE , the gels were analyzed with software ImageMaster4. 01 to find out the spots different between them, and the proteins in the spots were identified by matrix associated laser desorption Pionization time-of-flight (MALDI-TOF) mass spectrometry. Results Systemic processing and essential conditions of sample prepraration and 2-DE for transparent lens and opacity lens were established successfully.There were more than 500 spots with good distribution on the gel for each tissue.Decades of different spots were displayed between them. The maps of 2-DE showed that lens proteins that lens proteins were in the section of PH 4-5,and the relative molecularweight was 20000-30000, transparent lens and opacity lens was raised and candidate marker of one crystalline of CRYBA3/A1 were found under such strategy.Conclusion Under reasonable strategy , the analysis of proteomics with 2DE on human tissue is a useful method for discovering valuable disease marker candidates. CRYBA3/A1which were found in lens tissues will be very important in diagnosis and treatment of congenital nuclear cataract.
[ Key words] Proteomics；Congenital nuclear cataract；Tow-dimensional electrophoresis； Mass spectrometry；
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