HIF-1α介导低氧诱导的内皮细胞Brm表达上调的机制研究

发布时间：2018-02-27 17:21

本文关键词： 低氧性肺动脉高压低氧诱导因子α Brm蛋白内皮细胞　出处：《中国病理生理杂志》2017年04期 　论文类型：期刊论文

【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。
[Abstract]:Aim: to investigate the role and mechanism of hypoxia inducible factor-1 伪 (HIF-1 伪) in the up-regulation of Brm expression in cultured vascular endothelial cells induced by hypoxia. Methods: overexpression of HIF-1 伪 in endothelial cells or siRNA interference with HIF-1 伪 expression were observed after treatment with 1% hypoxia for 24 h. The transcriptional activity of Brm promoter was detected by luciferase reporter gene assay, and the mRNA expression level of Brm was detected by RT-qPCR. Western blot was used to detect the expression of Brm protein, and ChIP technique was used to detect the binding changes of HIF-1 伪 and Brm promoter region under hypoxia. Results: overexpression of HIF-1 伪 could significantly up-regulate the expression of HIF-1 伪 and protein in Brm promoter. However, siRNA interference with HIF-1 伪 could significantly inhibit the expression of Brm promoter activity mRNA and protein. The experiment showed that hypoxia could significantly enhance the binding of HIF-1 伪 to Brm promoter. Conclusion: hypoxia upregulated the expression of Brm in endothelial cells dependent on the transcriptional regulation of HIF-1 伪.
【作者单位】：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;第三军医大学高原军事医学系全军高原医学重点实验室;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;
【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)
【分类号】：R544.1

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