成年人与2个月月龄婴儿视网膜的蛋白质组学初步研究

Human retinal proteomics study of adults and two-month old infants

目的 建立成年人与2个月月龄婴儿视网膜组织的双向凝胶电泳图谱,初步分析成年人视网膜与婴儿视网膜蛋白质的表达差异.方法 9例角膜移植后剩余的成年人眼球视网膜组织,2例角膜移植后剩余的2个月月龄婴儿眼球视网膜组织;抽提蛋白质,双向凝胶电泳后考马斯亮蓝染色扫描分析图谱,选取明显差异蛋白点进行质谱鉴定.结果 成年人与2个月月龄婴儿视网膜组织的凝胶经考马斯亮蓝染色分别检测到1179和1295个蛋白点,其中1039个蛋白点位置相匹配.5个差异蛋白点的质谱鉴定结果显示,人血清白蛋白、鸟苷酸激酶在成年人的视网膜中高表达;微管蛋白、转醛醇酶和晶体蛋白在2个月月龄婴儿的视网膜中高表达.结论 成功建立了人视网膜蛋白质的双向凝胶表达图谱.经质谱鉴定证实了成年人和婴儿的视网膜蛋白质存在表达差异.

Objective To identify proteins that have expressed in human eyes from adults and two-month old infants by proteomics approach, so as to build a two-dimensional gel electrophoresis （two-DE） reference map for human retina. The difference of proteomics between the retinas of adults and two-month old infants are also studied. Methods Human retina tissues were collected from donor eyes （nine adults and two infants）. Proteins were separated by two-DE. The gels were analyzed by image software. Protein spots were excised from the gels and detected by matrix assisted laser desorption ionization time off light mass spectrometry （MALDI-TOF-MS）. Results A total of 1179 spots and 1295 spots were detected respectively on two-DE gels of Coomassie-stained adults and two-month old infants retina, of which 1039 spots were matched in the position. Five spots up-regulated were successfully identified. Human serum albumin and 4 guanylate kinase 1 （GUK1） were identified in adult retina. β2-tubulin, transaldolasel and alpha A-crystallin were identified in infant retina. Conclusion The two-DE reference map for retina proteomics is successfully established. This study provides an evidence of changes in retinal protein levels between adults and infants and biochemical pathways for future studies of human retina development.