AP-1-dependent transcriptional regulation of NADPH oxidase in human aortic smooth muscle cells role of p22phox subunit
AP-1-dependent transcriptional regulation of NADPH oxidase in human aortic smooth muscle cells role of p22phox subunit
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DOI：
10.1161/atvbaha.108.163592
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发表时间：
2008-05-01
影响因子：
8.7
通讯作者：
Simionescu, Maya
中科院分区：
文献类型：
--
作者：
Manea, Adrian;Manea, Simona A.;Simionescu, Maya
目的：nadph氧化酶（NADPHox）是血管疾病中活性氧的主要来源；酶活化的机制尚未完全阐明。AP-1控制许多与血管平滑肌细胞（SMCs）功能障碍相关的基因表达。在这项研究中，我们寻找AP-1在促炎条件下对人主动脉SMCs中NADPHox表达和功能的调节作用。方法与结果：培养的SMCs分别暴露于血管紧张素II （Ang II）或肿瘤坏死因子(TNF)- α。荧光素增强的化学发光实验和实时聚合酶链反应分析显示，AP-1和丝裂原激活的蛋白激酶抑制剂降低了Ang II或tnf α依赖的NADPHox活性和mRNA表达上调(NOX1, NOX4, p67(phox), p47（phox), p22(phox)）。AP-1抑制剂显著降低Ang II或tnf - α刺激的p22（phox）启动子活性和蛋白水平。c-Jun/c-Fos的瞬时过表达上调了p22（phox）启动子活性。转录因子下拉实验和染色质免疫沉淀证实了c-Jun蛋白与p22（phox）基因启动子中预测的ap -1结合位点的物理相互作用。结论：在暴露于Ang II或tnf - α的SMCs中，抑制ap -1相关通路可降低NADPHox的表达和o -2的产生。AP-1与p22（phox）基因启动子的物理相互作用促进了NADPHox的调控。
Objective-NADPH oxidase (NADPHox) is the major source of reactive oxygen species in vascular diseases; the mechanisms of enzyme activation are not completely elucidated. AP-1 controls the expression of many genes linked to vascular smooth muscle cells (SMCs) dysfunction. In this study we searched for the role of AP-1 in the regulation of NADPHox expression and function in human aortic SMCs exposed to proinflammatory conditions.Methods and Results-Cultured SMCs were exposed to either angiotensin II (Ang II) or tumor necrosis factor (TNF)-alpha. The lucigenin-enhanced chemiluminescence assay and real-time polymerase chain reaction analysis revealed that AP-1 and mitogen-activated protein kinase inhibitors reduced both Ang II or TNF-alpha-dependent upregulation of NADPHox activity and mRNA expression (NOX1, NOX4, p67(phox), p47(phox), p22(phox)). Inhibitors of AP-1 significantly diminished the Ang II or TNF-alpha-stimulated p22(phox) promoter activity and protein level. Transient overexpression of c-Jun/c-Fos upregulated p22(phox) promoter activity. Transcription factor pull-down assay and chromatin immunoprecipitation demonstrated the physical interaction of c-Jun protein with predicted AP-1-binding sites in the p22(phox) gene promoter.Conclusions-In SMCs exposed to Ang II or TNF-alpha, inhibition of AP-1-related pathways reduces NADPHox expression and the O-2-production. The physical interaction of AP-1 with p22(phox) gene promoter facilitates NADPHox regulation.