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目的:建立分辨率高和重复性好的鼻息肉病及对照鼻黏膜双向凝胶电泳(2DE)图谱,筛选鉴定差异表达蛋白质。方法:收集鼻息肉病(鼻息肉病组)及对照鼻黏膜标本(对照组)各7例,固相pH梯度2DE、凝胶银染,扫描图像,ImageMaster2DEElite4.01软件分析,识别差异表达蛋白质,质谱仪得到相应肽质指纹图谱(PMF),PeptIdent软件查询SwissProtandTreMBL数据库,鉴定差异表达蛋白质。结果:①鼻息肉病组和对照组各自随机挑选的1个样本3次重复2DE,平均蛋白质点数分别为891±67和936±62;匹配点数为767±83和821±78,平均匹配百分率为86.1%和87.7%;同一鼻息肉病的3块胶在蛋白质点位置上有较好的重复性,不同胶间蛋白质点在等电聚焦电泳(IEF)方向偏差为(1.13±0.16)mm,在SDS聚丙烯酰胺凝胶电泳(SDSPAGE)方向偏差为(1.45±0.21)mm。分析2种组织各7例样本的2DE图谱的平均凝胶图谱,鼻息肉病组和对照组蛋白质点数为1532和1617,匹配点数为1065个。②质谱分析差异蛋白质点20个,获取16张PMF,查询数据库鉴定出差异蛋白质11个。结论:本研究建立了分辨率高和重复性好的鼻息肉病及鼻黏膜的2DE图谱,识别鉴定出一些与鼻息肉病变相关的差异表达蛋白质。
Objective: To establish a high resolution and reproducible nasal polyposis and control nasal mucosa two - dimensional gel electrophoresis (2DE) map, screening and identification of differentially expressed proteins. Methods: Seven cases of nasal polyposis (nasal polyposis disease) and control nasal mucosa (control group) were collected. Seven samples were collected and analyzed by 2D phase gel electrophoresis, gel silver staining, scanning image and ImageMaster2DEElite4.01 software to identify differentially expressed proteins, Mass spectrometry obtained corresponding peptide fingerprinting (PMF), PeptIdent software query SwissProtandTreMBL database to identify differentially expressed proteins. Results: ① One specimen randomly selected from nasal polyposis and control group was repeated 3 times with 2DE, the average number of protein spots were 891 ± 67 and 936 ± 62 respectively; the matching points were 767 ± 83 and 821 ± 78, the average matching percentage was 86.1% and 87.7%, respectively. The three gels of the same polyposis disease had good repeatability at the protein spots. The protein spots in different gels had (1.13 ± 0.16) mm in the direction of isoelectric focusing (IEF) SDS polyacrylamide gel electrophoresis (SDSPAGE) direction deviation (1.45 ± 0.21) mm. The average gelatinograms of 2DE images of seven samples from two tissues were analyzed. The number of protein spots in nasal polyposis and control groups was 1532 and 1617, and the matching points were 1065. ② Mass spectrometry analysis of differential protein spots 20, access to 16 PMF, query database identified 11 differential proteins. Conclusion: This study established a high resolution and good reproducibility of nasal polyposis and nasal mucosa of the 2DE pattern, identification identified some of the nasal polyp lesions associated with the differential expression of proteins.