2型糖尿病患者泪液蛋白表达分析

Expressional analysis of tear proteins in type 2 diabetic patients

目的 通过蛋白质组学技术分析2型糖尿病患者泪液中蛋白质的表达.方法 选取2007年2月至2008年2月西安交通大学医学院第一附属医院内分泌科住院的2型糖尿病合并视网膜病变患者15例(DR组)、无视网膜病变患者15例(NDR组)、非糖尿病正常对照者15名(CTL组),收集3组研究对象的基础泪液约5～10μl,采用双向凝胶电泳技术和Progenesis Samespots图像分析软件对3组泪液中表达差异的蛋白质进行分析.采用单因素方差分析比较3组间蛋白点表达的差异,对有差异的变量采用Bonferroni检验进行两两比较.各蛋白点表达水平与糖化血红蛋白(HbAlc)、糖尿病病程间的关系采用Pearson相关性分析.结果 与CTL组相比,DR组11种泪液蛋白表达上调(F值分别为3.635、3.479、4.427、5.031、3.508、5.253、3.368、4.186、3.669、3.778和3.856,均P＜0.05),DR组、NDR组患者5种泪液蛋白的表达下调(F值分别为5.149、8.179、6.692、4.733和3.539,均P＜0.05);另外,DR组患者6种泪液蛋白的表达显著高于NDR组患者(F值分别为4.427、5.031、4.186、3.669、3.778和3.856,均P＜0.05).糖尿病患者泪液蛋白Spot200的表达水平与HbAlc水平呈显著正相关(r=0.370,P＜0.05),其他15种泪液蛋白(Spot2、32、251、180、167、 199、231、185、283、80、197、132、137、242、255)的表达水平与HbAlc之间无相关性显示.另外,蛋白点的表达水平与糖尿病病程无显著相关性(均P＞0.05).结论 糖尿病视网膜病变可以导致泪液中蛋白质表达水平变化,这些蛋白表达对理解该病的发病机制、评估治疗具有重要的价值.

Objective To investigate the expression of tear proteins in type 2 diabetic patients by proteomics technique. Methods Fifteen type 2 diabetic patients without retinopathy, 15 patients with retinopathy, and 15 age and gender-matched non-diabetic controls were recruited and 5 - 10 μl basal tear samples were collected. The expressional difference of tear proteins among the three groups were analysed with 2-DE and Progenesis Samespots software. The statistical difference among the three groups were estimated by one-way analysis of variance followed by Bonferroni multiple comparison test. Pearson correlation analysis were performed between expressional volume of tear proteins and HbAlc, diabetic duration. Results In comparison with non-diabetic controls, 11 tear protein expressions in diabetic patients with retinopathy were up-regulated (F value was 3. 635, 3.479, 4. 427, 5.031, 3. 508, 5. 253, 3. 368,4. 186, 3.669, 3. 778, and 3. 856, respectively, all P＜0.05), while 5 proteins down-regulated in diabetic patients with and without retinopathy ( F value was 5. 149, 8. 179, 6. 692, 4. 733, and 3. 539, respectively,all P＜0.05). In addition, the expressional levels of 6 tear proteins were higher in diabetic patients with retinopathy than those without retinopathy ( F value was 4. 427, 5. 031,4. 186, 3. 669, 3. 778, and 3. 856,respectively, all P ＜ 0. 05 ). A significant correlation was found between the expressional volume of spot 200and HbA1c(r =0. 370,P＜0.05)in diabetic patients, but no any correlation was demonstrated between HbA1c and expressional volumes of the other 15 spots, or between diabetic duration and expressional volumes of spots (all P ＞ 0. 05 ). Conclusions These differentially expressed tear proteins in diabetic retinopathy might be useful in understanding the pathogenesis and evaluating treatment.