摘要
目的探讨肿瘤坏死因子(TNFα)通过其Ⅱ型受体介导的细胞内信号机制。方法采用TNFαⅠ型受体敲除的脑血管内皮细胞[BVEC/TNF-RI(-/-)]为研究对象,用氧化-还原敏感的荧光探针2,7-Dichlo-rofluorescein(DCF)检测细胞内氧化-还原水平,用western blot法检测TNFα对JNK、ERK激酶和转录因子AP-1活性的影响,并用小分子抗氧化剂NAC或JNK、ERK抑制剂干预上述诱导实验。结果TNFα(5ng/mL)处理24h,BVEC/TNF-RI(-/-)内活性氧产生增多(P<0.05)。此外,TNFα刺激BVEC/TNF-RI(-/-)可引起JNK和ERK激酶表达和转录因子AP-1的活性增强(P<0.05),JNK的抑制剂SP600125能减弱TNFα诱导的AP-1的表达(P<0.05),而NAC、ERK的抑制剂不能。结论TNFα可通过其Ⅱ型受体介导细胞内活性氧的产生,JNK、ERK激酶的磷酸化和转录因子AP-1的激活。
[Objective] To investigate the intracellular signaling mechanisms mediated by Tumor Necrosis Factor α（TNFα） Receptor Ⅱ. [Methods] TNFα receptor Ⅰ （TNFR1） knockout murine brain microvessel endothelial cells （BMVECs） in vitro culture were stimulated respectively by TNFα （5 ng/mL）, The oxidative stress was measured in the cells by using 2, 7-Dichlorofluor-escein（DCF） diacetate as probe,The effect of TNFα on JNK, ERK or Transcription factor AP-1 were analyzed by Western blot, while micromolecule antioxidant NAC, ERK inhibitor（PD98059） or JNK inhibitor （SP600125） were used. [Results] Exposure of TNFα receptor Ⅰ （TNFR1） knockout BMVECs [BVEC/ TNF-RI（-/-）] to TNFα （5 ng/mL, 24 h） caused a marked increase in the level of reactive oxygen species（ROS）. Furthermore, TNFα stimulation also lead to the activation of JNK,ERK and the Transcription Factor AP-1 in the BVEC/TNF-RI（-/-）. However, incubation with the JNK inhibitor SP600125 but not the ERK inhibitor PD98095 or NAC attenuated the TNFα-induction of AP-1 in the BVEC/FNF-RI（-/-）. [Conclusion] TNFα receptor Ⅱ may play a major role in the TNFα-induced the increase of ROS, phosphorylation of JNK or ERK,and the activation of Transcription Factor AP-1.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2008年第15期2127-2130,共4页
China Journal of Modern Medicine
基金
黑龙江省自然科学基金资助(No:ZA2006-05)
黑龙江省教育厅科学技术研究项目(No:11521328)
关键词
肿瘤坏死因子Ⅱ型受体
信号传导
Tumor Nesrosis Factor Receptor Ⅱ
signal transduction
