常见舌苔蛋白质组学与生物信息学研究

Study on Proteomics and Bioinformatics of the Common Tongue Coating

目的运用蛋白质组学技术探讨不同舌苔的差异表达蛋白质。方法将观察对象根据有无各系统器质性疾病及舌苔表现,分为正常薄苔组、病理薄苔组、厚苔组和剥苔组,以固相pH梯度等电聚焦为第一相、垂直SDS-PAGE为第二相进行双向电泳,银染显色,计算机扫描,Image Master2D Platinum软件分析;选择差异点胶内胰酶消化后,进行MALDL-TOF-MS蛋白质质谱鉴定和生物信息学分析。结果四组舌苔组织可检测的蛋白质斑点数分别为(1 082±105)个、(1 052±85)个、(1 129±98)个、(1 143±140)个。差异蛋白质点肽质指纹的质谱鉴定和分析生物信息学分析结果,鉴定出13个舌苔密切相关蛋白质,其中cystatin B、cytokeratin 13和GAL7 protein为首次发现的舌苔变化相关蛋白,与舌苔发生发展的关系密切;并发现了5个新的舌苔相关未知蛋白,在目前国际蛋白质数据库中无收载,有必要进一步深入研究。结论不同病理舌苔与正常舌苔的蛋白质表达谱存在明显差异。

Objective To investigate differential expression proteins among different tongue coatings with protecomics.Methods The objects in observation were divided into normal thin white tongue coating group,pathological thin tongue coating group,thick tongue coating group and pelt tongue coating group according to whether suffering from organic diseases and with the pathological tongue coatings.Two-dimensional electrophoresis（2DE）was conducted,in which,the immobilized pH gradient isoelectric focusing was taken as the first dimension and the vertical SDS-PAGE as the second one.Silver staining coloration,CT scanning,ImageMaster 2D Platinum software analysis were applied.After glue trypsin digestion of the selected different points,MALDL-TOF-MS protein mass spectrometry and bioinformatics analysis were performed.Results The numbers of protein spots detected in tongue coating tissue of four groups were（1 082±105）,（1 052±85）,（1 129±98）and（1 143±140）separately.With mass spectrometry of peptide mass fingerprinting（PMF）of differential protein spots and bioinformatics analysis,it was identified that 13 proteins were closely relevant with tongue coating,of which,cystatin B,cytokeratin 13 and GAL7 protein were the proteins discovered at the first time in the relation with the changes in tongue coating,indicating tight connection with the occurrence and development of tongue coating.Additionally,5 unknown proteins were newly discovered to be relevant with tongue coating,which had not been collected in the international protein database at present yet and it was necessary to have a further study on their biological significance.Conclusion There are obvious differences in the protein expression profiling between pathological tongue coatings and normal tongue coating.