鼻息肉、鼻息肉病及对照鼻黏膜蛋白质双向电泳图谱的建立

Establishment of two-dimensional electrophoresis profiles of proteins from human nasal polyps and polyposis and nasal mucosa

目的 建立鼻息肉、鼻息肉病及对照鼻黏膜组织蛋白质双向凝胶电泳 (2 DE)图谱。方法 收集并- 80℃冻存鼻息肉、鼻息肉病及鼻黏膜组织样本各 3例 ,双向凝胶电泳分离样本总蛋白、银染凝胶显色 ,扫描图像 ,ImageMaster 2 DEElite4 .0 1软件分析。结果 鼻息肉、鼻息肉病和鼻黏膜组织同一样本三块凝胶平均蛋白质点数分别为 82 5± 78个 ,891± 6 7个和 936± 6 2个 ;平均匹配点数分别为 6 82± 96个 ,76 7± 83个和 82 1± 78个 ,其平均匹配百分率分别为 82 .7%、86 .1%和 87.7% ;位置重复性分析 ,IEF方向平均偏差为 1.13± 0 .16mm ,SDS PAGE方向平均偏差为 1.4 5± 0 .2 1mm。鼻息肉、鼻息病和鼻黏膜三种组织各 3例样品的平均蛋白质点数分别为 876± 95个 ,95 3± 84个和 10 16± 79个。三种组织的 2 DE平均凝胶图谱 :鼻息肉、鼻息肉病和鼻黏膜蛋白质点数分别为116 2个、12 74和 1336个 ,平均匹配点数为鼻息肉与鼻黏膜之间 75 7个 ;鼻息肉病与鼻黏膜之间为 82 5个 ;鼻息肉与鼻息肉病之间为 883个。结论 本实验建立了图像清晰、分辨率高和重复性好的鼻息肉、鼻息肉病及鼻黏膜组织的固相PH梯度 2 DE图谱 ,有助于进一步识别鉴定三者差异表达的蛋白质。

Objective To establish two-dimensional gel electrophoresis (2-DE) of proteins from human nasal polyps and polyposis and nasal mucosa. Methods Samples of human nasal polyps, polyposis and nasal mucosa tissues ,each tissue sample contained 3 cases obtained, snap-frozen in liquid nitrogen immediately and then stored at -80℃. The total proteins were extracted and separated by 2-DE. The silver-stained 2-DE were scanned with digital ImageScanner and analyzed with ImageMaster 2-DE Elite 4.01 software. Results 2-DE for the 3 tissues from the same patient was repeated 3 times, respectively. The image analyasis showed: For the polyps tissues, the average proteins spots of 3 gel were 825±78 and 682±96 spots were matched with the average matching rate of 82.7%; For the polyposis tissues, the average proteins spots of 3 gel were 891±67 and 767±83 spots were matched with the average matching rate of 86.1%; For the nasal mucosa tissues, the average proteins spots of 3 gel were 936±62 and 821±78 spots were matched with the average matching rate of 87.7%. The average deviations of matched spot position were 1.13± 0.16 mm in IEF direction and 1.45± 0.21 mm in SDS-PAGE direction, respectively. Analysising the 2-DE profiles of 3 tissues, each tissue were cotaining 3 cases obtained, the average proteins spots of nasal polyps, polysis and mucosa were 876±95, 953±84 and 1016±79. Comparing the average electrophoresis profiles of the 3 tissues. The proteins spots of nasal polyps, polyposis and nasal mucosa tissues were 1162, 1274 and 1336. The matched proteins spots of the average electrophoresis profiles of the three tissues: For the nasal polyps and nasal mucosa, were 757; For the nasal polypsis and nasal mucosa, were 825; For the nasal polyps and nasal polypsis, were 883 respectively. Conclusion In this study, the well-resolved, reproducible 2-DE patterns of human nasal polyps and polyposis and nasal mucosa were established. It is helpful to further identify the differential proteins of the three tissues.