论文部分内容阅读
目的应用双向电泳及质谱技术对野生型及变异型白细胞介素13(IL-13)刺激后的ASMC 总蛋白进行差异比较研究,借此分析差异表达蛋白与支气管哮喘的关系,为哮喘的治疗提供新的靶标。方法用固相 pH 双向电泳分离,分别给予野生型 IL-13及变异型 IL-13刺激的 ASMC 总蛋白组成分,找出异常 IL-13处理后与野生型 IL-13处理后有明显差别的蛋白斑点,用基质辅助激光解吸电离飞行时间串联质谱(MALDI-TOF)进行鉴定部分差异蛋白质。结果获得了分辨率高,重复性好的电泳图谱,在野生型 IL-13刺激组和变异型 IL-13刺激组分别平均有(840±21)个和(892±17)个蛋白点(n=3)被检测,匹配的点数为(685±19)个,选取5个未匹配点做质谱分析,鉴定其中3个分别为 stathmin 1,Ribosomal protein p0,NADH dehydrogenase。结论本研究结果显示不同处理组的ASMC 总蛋白表达存在差异,这些差异蛋白有可能为阐明野生型 IL-13及其变异体引发哮喘机制提供新的实验证据。
OBJECTIVE: To compare the differences in the total proteins of ASMCs induced by wild-type and variant interleukin-13 (IL-13) using two-dimensional electrophoresis and mass spectrometry to analyze the relationship between differentially expressed proteins and bronchial asthma and to provide the treatment of asthma New target Methods Two-dimensional gel electrophoresis was used to separate the ASMCs from wild-type IL-13 and mutant IL-13. The results showed that there were significant differences between the two groups Protein spots were identified using a matrix-assisted laser desorption / ionization time-of-flight tandem mass spectrometry (MALDI-TOF) for partial differential protein identification. RESULTS: The electropherogram with high resolution and good reproducibility obtained an average of (840 ± 21) and (892 ± 17) protein spots in the wild-type IL-13 stimulation group and the mutant IL- = 3) were detected, the number of matching points was (685 ± 19). Five unmatched spots were selected for mass spectrometry analysis. Three of them were identified as stathmin 1, Ribosomal protein p0 and NADH dehydrogenase respectively. Conclusion The results of this study show that there are differences in the total protein expression of ASMC in different treatment groups. These differential proteins may provide new experimental evidence for elucidating the mechanism of asthma induced by wild-type IL-13 and its variants.