慢性肾小球肾炎肾阴虚证血浆蛋白质双向凝胶电泳分析

Two-dimensional gel electrophoresis analysis on plasma proteomics of chronic glomerulonephritis of Kidney-YIN deficiency syndrome

目的初步观察慢性肾小球肾炎肾阴虚证患者血浆蛋白质组的变化与表达差异,为进一步探讨肾阴虚证的发生机制奠定基础。方法采用双向凝胶电泳(2-DE)技术分离慢性肾小球肾炎肾阴虚证患者、肾阳虚证患者和正常人的血浆总蛋白,银染显色;PDQest软件对凝胶图像进行定性定量差异表达分析;从中选取差异表达蛋白质斑点,通过基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)分析,获取肽质量指纹图谱(PMF);通过Mscot-Wizard软件进行数据库搜索,鉴定蛋白质。结果与正常人和慢性肾小球肾炎肾阳虚证患者相比,在慢性肾小球肾炎肾阴虚证患者血浆蛋白质双向凝胶电泳图谱中绝对高表达的蛋白质斑点有80个,绝对低表达的蛋白质斑点有42个;定性差异表达分析发现,在慢性肾小球肾炎肾阴虚证患者血浆蛋白质双向凝胶电泳图谱中特异出现的蛋白质斑点有2个,缺失的蛋白质斑点有13个。结论同病异证患者血浆中存在差异表达蛋白质,利用蛋白质组学技术有望对中医证候的发生机制进行阐释。

Objective To observe the differential expression of plasma proteomic in chronic glomerulonephritis of Kidney-YIN deficiency syndrome patient and to establish the basis for further research.Methods Total proteins in plasma of chronic glomerulonephritis Kidney-YIN deficiency syndrome,Kidney-YANG deficiency syndrome patients,and normal subjects were studied by two-dimensional gel electrophoresis（2-DE） isolation technique and silver staining methods.The qualitative and quantitative expression differences were analyzed by PDQest gel image software.Peptide mass fingerprinting（PMF）of the differentially expressed protein spots were analyzed by matrix-assisted laser desorption ionization time of flight mass spectrometry（MALDI-TOF-MS）.The protein database was searched by Mscot-Wizard software to further characterize the differential protein expression.Results Compared with the normal subjects and chronic glomerulonephritis Kidney-YANG deficiency syndrome patients,there were 80 highly expressed protein spots and 42 lowly expressed protein spots in the 2-DE images of Kidney-YIN deficiency syndrome patients.According the qualitative analysis of differential expression,there were 2 unique protein spots and 13 missing protein spots on the 2-DE images of Kidney-YIN deficiency syndrome patients.Conclusion Differential plasma protein expression was noted in patients with the same disease and different syndromes.Proteomics technology may be used to study the mechanism of traditional chinese medicine（TCM） syndromes.