【摘要】 目的:探讨在炎症刺激因子肿瘤坏死因子-α(TNF-α)介导的炎症反应中转录因子GATA家族成员是否参与了Tie2基因的转录调控。方法:采用实时定量PCR测定TNF-α作用前后人主动脉内皮细胞(HAECs)和肺动脉内皮细胞(HPAECs)中GATA家族成员GATA2、GATA3 mRNA的表达,利用荧光素酶活性检测研究GATA转录因子能否激活Tie2基因启动子,并采用电泳迁移变动分析(EMSAs)和超级迁移率变动试验(super-shift)研究GATA转录因子是否通过与Tie2基因启动子结合,从而激活Tie2基因的表达。结果:TNF-α可诱导人血管内皮细胞HAECs和HPAECs中转录因子GATA2的表达,高表达的GATA2可与Tie2基因启动子上的(T/A)GATA(A/G)序列结合,激活Tie2基因启动子,从而上调Tie2基因的表达。结论:GATA转录因子家族成员GATA2在TNF-α介导的炎症反应中调节了Tie2基因的表达。更多还原

【Abstract】 AIM: To explore the possibility that the transcription factor GATA family member is involved in the regulation of Tie2 gene expression in the tumor necrosis factor-α(TNF-α) mediated inflammatory response.METHODS: The mRNA expression of GATA family members GATA2 and GATA3 was determined in primary human aortic endothelial cells(HAECs) and primary human pulmonary artery endothelial cells(HPAECs) before and after TNF-α treatment by real-time quantitative PCR.The activation of Tie2 gene promoter by GATA transcription factor was detected by luciferase assay.The electrophoretic mobility shift assays(EMSAs) and supershift assay were performed for verifying whether Tie2 gene expression was activated by the binding of GATA factors to Tie2 promoter.RESULTS: TNF-α increased the expression of GATA2 in HAECs and HPAECs.The highly expressed GATA2 was bound to the sequence(T/A) GATA(A/G) of Tie2 promoter,activated Tie2 gene promoter and up-regulated Tie2 gene expression.CONCLUSION: GATA transcription factor family member GATA2 up-regulates the expression of Tie2 gene in TNF-α mediated inflammatory response.更多还原