TNF-α诱导类风湿关节炎滑膜成纤维细胞MAPKs通路的活化

Activation of MAPKs in signal transduction of TNF-α in fibroblast-like synoviocytes of rheumatoid arthritis

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

Objective:To study on MAPKs activation in rheumatoid arthritis fibroblastlike synoviocytes (RA FLS) under the stimulation of TNFα.Methods:RA FLS were primarily cultured;Western blot was applied to examine transient change of protein tyrosine phosphoralation status and MAPKs activation in RA FLS stimulated with various doses and periods of TNFα.Results:TNFα transiently increased protein tyrosine phosphoralation,and activated MAPKs cascades(mainly ERK 2,JNK 2,P 38 ) in RA FLS.The peak activation of ERK 2 and JNK were abstained at 10 U/ml,but that of P 38 was at 100 U/ml.The maximal activations of ERK 2,JNK 2 and P 38 took place at 5 min、15 min and 15 min respectively after stimulation with TNFα.Conclusion:During signal transduction of TNFα in RA FLS,tyrosine phoshorylation were increased transiently,MAPKs cascades were activated in a few minutes and there was heterogenicity of activation among three subfamily members.