论文部分内容阅读
目的 :研究INF α在类风湿关节炎 (Rheumatoidarthritis,RA)成纤维样滑膜细胞 (Fibroblast likesynoviocyte ,FLS)信号转导中MAPKs激活情况。方法 :原代培养类风湿性关节炎滑膜成纤维细胞 ;应用Westernblot检测TNF α短时间内引起RAFLS蛋白质酪氨酸磷酸化状态变化 ,及其对MAPKs家族成员活化的浓度效应和时相特点。结果 :INF α可以瞬时引起RAFLS蛋白质酪氨酸磷酸化程度增加 ;并在短时间内激活MAPKs通路 (ERK2 、JNK2 、P38为主 ) ,不同浓度梯度TNF α作用显示 :10U/ml时对ERK2 、JNK2 即可达到峰值活化 ,10 0U/ml时P38达到最大活化。时间上 ,ERK2 、JNK2 、P38的活化分别在TNF α作用后 5min、15min、15min最明显。结论 :INF α在RAFLS信号转导中 ,可以瞬时导致蛋白质酪氨酸磷酸化程度增加 ,并同时激活MAPKs 3条通路 ,但是MAPKs 3个亚家族成员的活化具有异质性
AIM: To investigate the activation of MAPKs in INFb signaling in rheumatoid arthritis (RA) fibroblast synovial cells (FLS). Methods: Primary cultured rheumatoid arthritis synovial fibroblasts were cultured in vitro. The changes of tyrosine phosphorylation status of RAFLS protein induced by TNFα and the concentration-effect and time-phase characteristics of MAPKs family member activation were detected by Western blot. Results: INFα could transiently induce the increase of tyrosine phosphorylation of RAFLS protein and activate MAPKs pathway in a short time (ERK2, JNK2 and P38). The effects of different concentrations of TNFα showed that ERK2, JNK2 peak activation can be reached, P10 10U / ml to achieve maximum activation. Time, ERK2, JNK2, P38 activation were TNFα after 5min, 15min, 15min most obvious. CONCLUSION: INF α can transiently lead to the increase of protein tyrosine phosphorylation in RAFLS signal transduction and activate the three MAPKs pathways simultaneously, but the activation of three subfamilies of MAPKs is heterogeneous