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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。
Objective: To investigate the protein composition of 11000 differential (abnormal) bands in thymus hyperplasia in patients with myasthenia gravis (MG). Methods: 11000 bands were screened and enriched by non-reducing SDS-PAGE preparative electrophoresis. Two-dimensional electrophoresis was used to obtain the two-dimensional map of proteins and the differential protein spots were analyzed by mass spectrometry. 4.1R protein expression in 10 cases of hyperplasia and 10 cases of normal control. Results: The positive rate of target band in thymus hyperplasia was 72.7% (8/11) in MG patients. The band consisted of (30 ± 6) protein spots, and there were 16 spots with high expression difference in mass spectrometry Three proteins were identified as 4.1R protein, self-assembly inhibitor 1 and choline acetyltransferase. The results of immunohistochemistry showed that the expression level of 4.1R protein in thymus hyperplasia MG group was (1.55 ± 0.46) (t = 3.054, P = 0.016) significantly higher than that in normal control group (0.98 ± 0.18). CONCLUSION: The 11,000 bands of MG thymus hyperplasia consist of multiple protein spots. The major functions of these proteins are involved in the composition of cytoskeleton, intracellular information transmission and acetylcholine synthesis.