HIF-1α介导低氧诱导的内皮细胞Brm表达上调的机制研究  

HIF-1α mediates hypoxia-induced Brm transactivation in endothelial cells

作　　者：王授衔 陈德伟[1,2] 高文祥[1,2] 徐刚[3,2] 吴刚[1,2] 陈建[3,2] 高钰琪[3,2] 

机构地区：[1]第三军医大学高原军事医学系病理生理学与高原病理学教研室,重庆400038 [2]第三军医大学高原军事医学系全军高原医学重点实验室,重庆400038 [3]第三军医大学高原军事医学系高原特需药品与卫生装备研究室,重庆400038

出　　处：《中国病理生理杂志》2017年第4期577-582,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81501626);军队重大项目(No.AWS14C007);国家科技部"973"项目(No.2012CB518201)

摘　　要：目的:本研究拟探讨低氧诱导因子1α(HIF-1α)在低氧诱导的血管内皮细胞Brm表达上调中的作用及机制,为低氧性肺动脉高压的防治提供理论依据。方法:在内皮细胞中过表达或siRNA干扰HIF-1α的表达,并给予1%低氧处理24 h后,运用萤光素酶报告基因检测方法检测Brm启动子的转录活性,运用RT-qPCR检测Brm的mRNA表达水平,运用Western blot检测Brm的蛋白表达水平。运用ChIP技术检测低氧条件下HIF-1α与Brm启动子区域的结合变化情况。结果:过表达HIF-1α可显著上调Brm的启动子活性、mRNA及蛋白表达,而siRNA干扰HIF-1α可显著抑制Brm的启动子活性、mRNA及蛋白表达,ChIP实验揭示低氧可显著增强HIF-1α与Brm启动子的结合。结论:低氧上调内皮细胞中Brm的表达依赖HIF-1α的转录调控作用。AIM： To clarify the molecular mechanism of Brm activation in endothelial cells during hypoxia and hypoxic pulmonary hypertension. METHODS： Hypoxia-inducible factor 1 alpha（ HIF-1α） was over-expressed or depleted in the endothelial cells under hypoxia. Luciferase activity was assayed after transfection using a luciferase reporter assay system. The expression of Brm at mRNA and protein levels was detected by RT-qPCR and Western blot. The occupancy of HIF-1α on Brm promoter was detected by the method of chromatin immunoprecipitation（ ChIP）. RESULTS： Brm expression was induced in cultured endothelial cells challenged with hypoxia. Over-expression of HIF-1α enhanced,while the depletion of HIF-1α attenuated Brm transactivation and expression under hypoxia. The results of ChIP revealed that hypoxia up-regulated the occupancy of HIF-1α on Brm promoter. CONCLUSION： HIF-1α,which is induced by hypoxia,binds to Brm promoter,and further promotes Brm transactivation in endothelial cells.

关 键 词：低氧性肺动脉高压 低氧诱导因子1Α Brm蛋白 内皮细胞 

分 类 号：R392.12[医药卫生—免疫学]