病理性近视眼患者血清蛋白质组学分子标志物筛选  

Proteomics analysis of serum biomarks in patients with pathological myopia

作　　者：邵珺[1] 辛瑜[2] 李荣秀[3] 陈凤娥[1] 张皙[1] 许迅[1] 孙晓东[1] 樊莹[1] 

机构地区：[1]上海交通大学附属第一人民医院眼科,200080 [2]江南大学生物工程学院 [3]上海交通大学生物制造实验室

出　　处：《中华眼科杂志》2012年第3期246-252,共7页Chinese Journal of Ophthalmology

基　　金：国家重点基础研究发展（973）计划子课题（2011CB707506）;上海市眼底病重点实验室开放课题基金（07Z22911）

摘　　要：目的探讨血清样本中差异蛋白质含量异常升高与病理性近视眼发病的关系。方法病例对照研究。选取病理性近视眼患者和同期正常人各30例。将病理性近视眼患者分为黄斑中心凹区视网膜脱离组8例，视网膜地图状萎缩组5例，黄斑裂孔组11例，脉络膜新生血管组6例。采用正常人血清免疫家兔制备多克隆抗体，ProteinA柱纯化混合抗体，溴化氰活化琼脂糖凝胶激活并合成亲和介质，基于抗原、抗体吸附原理，取备用病理性近视眼患者及正常对照者各血清样本上PBS平衡过的抗体柱，去除大量背景蛋白；应用毛细管高效液相色谱和质谱法检测并分析剩余蛋白。差异蛋白质间的相关性采用非参数相关分析的K-S秩和检验。结果人选病理性近视眼患者血清样本去除大量背景蛋白后，分析并检测出4种表达明显增高的差异蛋白质：转甲状腺素蛋白（TTR）表达阳性18例（60．0％），结合珠蛋白（HP）表达阳性11例（36．7％），血清结合素（HPX）表达阳性10例（33．3％），载脂蛋白（APO）表达阳性8例（26．7％）；这4种差异蛋白质间的表达阳性率呈正相关性（TTR与HP、HPX、APO相关性：r=0．480，0．577，0．492；HP与TTR、HPX、APO相关性：r=0．480，0．783，0．636；HPX与TTR、HP、APO相关性：r=0．577，0．783，0．853；APO与，TTR、HP、HPX相关性：r=0．492，0．636，0．853；P〈0．05）。在黄斑裂孔组中，有7例TTR表达阳性；脉络膜新生血管组中，有6例患者HP、TTR表达均阳性，5例患者HPX表达阳性。结论血清蛋白质组学分子标志物筛选法可以快速、有效去除正常人群和病理性近视眼患者共有的蛋白，从而发现可能与病理性近视眼相关的蛋白。这些相关的功能蛋白有可能成为筛选病理性近视眼的分子标志物。Objective To identify the correlation between serum-based differentially expressed proteins and pathological myopia. Methods It was a case-control study. The serum of 30 pathological myopia patients and 30 age- and gender-matched normal controls were collected from Shanghai First People＇s Hospital affiliated to Shanghai Jiaotong University. These patients were divided into 4 groups including macular-off retinal detachment （8 cases）, retinal geographic atrophy （5 cases）, macular hole （ 11 cases） and choroidal neovascular （6 cases）. The serum specimens of normal controls were used as immunogen to immune rabbits in order to prepare polyclonal antibodies. Purified by Protein A cartridge, these mixed antibodies were then combined with CNBr-activated Sepharose so as to synthesize affinity medium which was finally used to treat the serum specimens. According to the theory of antigen and antibody, common background proteins would be deleted. The remaining non-binding proteins were analyzed by capillary high- performance liquid chromatography and LTQ-MASS. Nonparametric statistical analysis was used to detect the correlation between each differentially expressed protein. Results The result of HPLC and LTQ-MASS in 30 specimens of patients revealed 4 peaks of differentially expressed proteins including TTR （ positive in 18 specimens, 60% ） , HP（ positive in 11 specimens, 36.7% ） , HPX （ positive in 10 specimens, 33.3% ） , APO（positive in 8 specimens, 26. 7% ）. There were positive correlations between these 4 proteins （the correlation between TTR and HP, HPX, APO is r = 0. 480, 0. 577, 0. 492 ; the correlation between HP and TTR, HPX, APO is r = 0. 480, 0. 783, 0. 636; the correlation between HPX and TTR, HP, APO is r = 0. 577, 0. 783,0. 853 ; the correlation between APO and TFR, HP, HPX is r =0. 492, 0. 636, 0. 853 ; P 〈 0.05 ）. In group of macular hole, TTR was positive expressed in 7 specimens while other differential proteins were low expression. In group of choroidal neovascular,

关 键 词：近视 退行性 蛋白质组学 生物学标记 前白蛋白 触珠蛋白类 血液结 合素 载脂蛋白B100 

分 类 号：R778.11[医药卫生—眼科]