基于2D-DIGE技术对早期糖尿病肾病肾阴虚证型患者血浆蛋白质组学研究

Research on Plasma Proteomics of Kidney Yin Deficiency of Early Diabetic Nephropathy Patients Plasma Proteomic Analysis Based on 2D-DIGE Technology

目的:寻找糖尿病肾病肾阴虚证敏感的血浆分子标志物。方法:对12例早期糖尿病肾病肾阴虚患者的血浆和12例正常血浆进行荧光差异双向电泳(2D-DIGE)分析,选择表达差异大于1.5倍的蛋白斑点进行质谱分析。结果:成功建立糖尿病肾病肾阴虚证和正常血浆的胶内差异双向凝胶电泳图谱,通过分析共鉴定出11种差异蛋白质,包括结合珠蛋白、凝溶胶蛋白、载脂蛋白AI等。结论:荧光差异双向电泳能够客观显示糖尿病肾病肾阴虚证与正常血浆之间的蛋白质表达差异,本研究鉴定的11种蛋白有可能为糖尿病肾病的早期诊断和中医证型研究提供潜在的血浆分子标志物。

Objective： To investigate the sensitive blood plasma molecular markers in kidney yin deficiency of diabetic nephropathy（ DN）. Methods： Two-dimensional fluorescence difference gel electrophoresis（ 2D-DIGE） was used to analyse the plasma which was from the patients with kidney yin deficiency of early diabetic nephropathy and the normal person from24 patients. Proteins that showed differential expression of a 1. 5- fold change were analyzed by MALDI-TOF / TOF mass spectrometry. Results： Two-dimensional electrophoresis（ 2-DE） maps of plasma proteins in patients with kidney yin deficiency of diabetic nephropathy（ DN） and healthy adults were established successfully. We validated 11 differentially expressed proteins detected by 2D,including HP protein、gelsolin、apolipoprotein A-I,etc. Conclusion： Proteomic analysis can objectively reveal the differences of protein expression between the two kinds of blood plasma. The 11 proteins that we have found have potential to provide plasma molecular markers for the clinical diagnosis and research on TCM pattern of diabetic nephropathy.