Liu Yin-Ping, Hu Shui-Wang, Wu Zhen-Feng, Mei Li-Xin, Lang Ping, Lu Xiao-He
Department of Ophthalmology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, Guangdong Province, China.
Int J Ophthalmol. 2011;4(6):616-22. doi: 10.3980/j.issn.2222-3959.2011.06.08. Epub 2011 Dec 18.
To establish and compare serum proteomic of diabetic retinopathy (DR) patients in various phases and discuss pathogenesis of DR so as to find out possible serum specific molecular markers for early diagnosis of DR.
Thirty-two subjects were divided into four groups: one group of eight type 2 diabetes mellitus (T2DM) patients without apparent DR (No-DR, NDR), one group of eight T2DM patients with non-proliferative diabetic retinopathy (NPDR), one group of eight T2DM patients with proliferative diabetic retinopathy (PDR) and one group of eight healthy volunteer participants. Two dimensional fluorescence difference gel electrophoresis (2D-DIGE) was applied to establish differential protein expression profiles in four groups. Matrix-assisted laser desorption/ionization time of flight tandem mass spectrometry (MALDI-TOF-TOF MS) was applied to identify mass spectrometry of differential proteins and analyze follow-up bioinformatics.
2D-DIGE maps of serum protein were satisfactory obtained from NDR, NPDR, PDR and normal control groups. Twenty-six different proteins spots were screened (the volume ratio was >1.5 based on DeCyder software analysis). Twenty-four of them were verified and two of them were not. Fifteen proteins were verified. Most of them were high-abundant proteins in serum. The four relatively low-abundant ones were beta 2-glycoprotein I (β(2)-GPI), alpha2-HS-glycoprotein(AHSG), alpha1-acid glycoprotein(α(1)-AGP) and apolipoprotein A-1(apo A-1). β(2)-GPI expression was gradually increased in the development of DR but unrelated to the severity of DR. The volume ratio of β(2)-GPI is 1.54, 2.43, and 2.84 in NDR, NPDR and PDR group respectively compared with normal control group.
Serum proteomic analysis of 2D-DIGE combined with MALDI-TOF-TOF MS is feasible to be applied in the study of DR. β(2)-GPI probably takes part in the process of DR occurrence and development and it could be a candidate biomarker on DR diagnosis in early phase.
建立并比较不同阶段糖尿病视网膜病变（DR）患者的血清蛋白质组，探讨DR的发病机制，以寻找可能用于DR早期诊断的血清特异性分子标志物。
32名受试者分为四组：一组为8名无明显DR的2型糖尿病（T2DM）患者（无DR，NDR），一组为8名患有非增殖性糖尿病视网膜病变（NPDR）的T2DM患者，一组为8名患有增殖性糖尿病视网膜病变（PDR）的T2DM患者，一组为8名健康志愿者。应用二维荧光差异凝胶电泳（2D-DIGE）建立四组的差异蛋白表达谱。应用基质辅助激光解吸/电离飞行时间串联质谱（MALDI-TOF-TOF MS）鉴定差异蛋白的质谱并分析后续生物信息学。
从NDR、NPDR、PDR和正常对照组成功获得血清蛋白的2D-DIGE图谱。筛选出26个不同的蛋白点（基于DeCyder软件分析，体积比>1.5）。其中24个得到验证，2个未得到验证。15种蛋白质得到验证。它们大多数是血清中的高丰度蛋白。四种相对低丰度的蛋白是β2-糖蛋白I（β(2)-GPI）、α2-HS-糖蛋白（AHSG）、α1-酸性糖蛋白（α(1)-AGP）和载脂蛋白A-1（apo A-1）。β(2)-GPI在DR发展过程中表达逐渐增加，但与DR的严重程度无关。与正常对照组相比，NDR、NPDR和PDR组中β(2)-GPI的体积比分别为1.54、2.43和2.84。
二维荧光差异凝胶电泳联合基质辅助激光解吸/电离飞行时间串联质谱的血清蛋白质组分析应用于DR研究是可行的。β(2)-GPI可能参与DR的发生发展过程，可能是DR早期诊断的候选生物标志物。
