重症肌无力患者胸腺增生组织中11000条带的蛋白质组学分析

Proteomics analysis of 11 000 protein band of myasthenia gravis patients with thymic hyperplasia

目的:探讨重症肌无力(MG)患者胸腺增生组织中相对分子质量为11000差异(异常)条带的蛋白质组成。方法:采用非还原性SDS-PAGE制备型电泳对11000条带进行筛选和富集;应用双向电泳得到蛋白质二维图谱,并对差异蛋白点进行质谱分析;应用免疫组化法比较20例MG胸腺增生组织和10例正常对照组织中4.1R蛋白的表达水平。结果:MG患者胸腺增生组织中目标条带的阳性率为72.7%(8/11),该条带由(30±6)个蛋白点组成,质谱分析显示有16个高表达差异蛋白点,其中3个蛋白得到鉴定,分别为4.1R蛋白、自整合抑制因子1和胆碱乙酰转移酶。免疫组化结果显示,与正常对照组的(0.98±0.18)相比,MG患者胸腺增生组织中4.1R蛋白表达水平为(1.55±0.46)明显增高(t=3.054,P=0.016)。结论:MG胸腺增生组织11000条带由多个蛋白点构成,这些蛋白主要功能涉及细胞骨架的组成、细胞内信息传递和乙酰胆碱合成等。

Aim：To discuss the formation and expression of abnormal 11 000 protein band from the tissue of myasthenia gravis patients（MG） with thymic hyperplasia.Methods：The 11 000 protein band was screened and enriched by non-reducing SDS-PAGE.The electrophoretogram was obtained from two dimensional electrophoresis,and differential proteins were analyzed by MS.The expression of protein 4.1R was compared between the experimental group（20 cases of MG） and control group（10 cases of normal control） using the method of immunohistochemistry.Results：The positive rate of 11 000 protein band was 72.7%（8/11）,which was consisted of（30±6）spots.16 protein spots expressed with higher level,among which,3 were identified as protein 4.1R,barrier to autointegration factor 1 and choline acetyltransferase,respectively.Immunohistochemical staining results showed that protein 4.1R from MG patients（1.55±0.46）was higher than that of control group（0.98±0.18）（t=3.054,P=0.016）.Conclusion：The 11 000 protein band is consisted of many protein spots,and main function of them is associated with the processes of cytoskeleton formation,signal transduction and acetylcholine synthesis.