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目的研究类风湿性关节炎(rheumatoidarthritis,RA)成纤维样滑膜细胞(fibroblast likesynoviocyte,FLS)信号转导中 ,IL 1β 介导的丝裂原活化的蛋白激酶(mitogen activated proteinkinaseMAPKs)的激活情况。方法原代培养RFFLS ;应用Westernblot检测IL 1β对于RAFLS蛋白质酪氨酸磷酸化状态的影响 ,及其对MAPKs家族成员活化的浓度效应和时相特点。结果IL 1β可以瞬时增加RAFLS蛋白质酪氨酸磷酸化的程度 ,并在短时间内激活MAPKs通路(以ERK2 ,JNK2和P38为主)。不同浓度的IL 1β对MAPKs的活化无明显差异 ,但ERK2,JNK2和P38的活化分别在IL 1β作用后5min ,15min和1min最明显。结论IL 1β在RAFLS信号转导中 ,可以瞬时导致蛋白质酪氨酸化的程度增加 ,并同时激活MAPKs3条通路 ;但MAPKs3个亚家族成员的活化具有异质性。
Objective To investigate the activation of IL 1β-mediated mitogen activated protein kinase (MAPK) activation in rheumatoid arthritis (RA) fibroblast likeynoviocyte (FLS) signal transduction. Methods The primary culture of RFFLS was performed. The effect of IL-1β on tyrosine phosphorylation status of RAFLS protein and the concentration-effect and time-phase characteristics of IL-1β on the activation of MAPKs family members were detected by Western blot. Results IL 1β transiently increased the level of tyrosine phosphorylation of RAFLS protein and activated MAPKs pathway in a short time (mainly ERK2, JNK2 and P38). However, the activation of ERK2, JNK2 and P38 were the most obvious at 5min, 15min and 1min after IL 1β treatment respectively. Conclusion IL-1β can transiently lead to the increase of protein tyrosination in RAFLS signal transduction and activate the MAPKs3 pathway at the same time. However, the activation of IL-1β in the three subfamilies of MAPKs is heterogeneous.