Control of activity and expression of ion transporters in rat lung: Role of HIF and CREB in hypoxia and beta-adrenergic stimulation

大鼠肺离子转运蛋白活性和表达的控制：HIF 和 CREB ​​在缺氧和 β-肾上腺素能刺激中的作用

• 批准号: 194361017
• 负责人: Professor Dr. Heimo Mairbäurl
• 金额: --
• 依托单位: Abteilung Innere Medizin VII: Sportmedizin
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2011
• 资助国家: 德国
• 起止时间: 2010-12-31 至 2015-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/194361017?language=en
• 关键词: Control; activity; expression; ion; transporters

ARDS, ALI, cardiovascular diseases, and alveolar hypoxia cause pulmonary edema, alveolar hypoxia, and impair protective mechanisms such as alveolar fluid reabsorption by decreasing activity and expression of transporters. It also induces inflammation, which further inhibits reabsorption. Protective mechanisms such as p2 adrenergic (ß2AR) signaling are impaired, which normally stimulates alveolar reabsorption by increasing expression and activity of ion transporters. Hypoxia-effects on gene expression are mediated by hypoxia-inducible factors (HIF), ß2AR-stimulation induces gene expression via CREB. Their role in the expression of ion transporters is unclear. Since both factors interact via co-activator proteins p300 and CBP, it is important to understand the interactions between effects of hypoxia and p2ARstimulation on alveolar reabsorption and expression of ion transporters, which is of great clinical relevance for treating pulmonary edema. In this project we want to test the hypotheses, that prolonged ß2AR stimulation can prevent the hypoxia-induced inhibition of activity and expression of alveolar ion transporters by CREB-dependent or -independent mechanisms. The roles of CREB and HIF and their interaction will be tested, and whether prolonged ß2AR stimulation prevents effects of hypoxia on alveolar transport. Experiments will be performed on primary rat and human alveolar epithelial cells invitro and on rats in-vivo exposed to hypoxia after treating with the ß2AR-agonist terbutaline for prolonged time to specifically evaluate the role of gene expression. The roles of HIF and CREB will be studied by gene silencing. Readout parameters are expression and activity of ion transporters and ß2AR-signaling.

ARDS、ALI、心血管疾病和肺泡缺氧导致肺水肿、肺泡缺氧，并通过降低转运蛋白的活性和表达损害保护机制，如肺泡液体重吸收。它还诱导炎症，进一步抑制重吸收。保护机制如β 2肾上腺素能（β 2AR）信号传导受损，其通常通过增加离子转运蛋白的表达和活性来刺激肺泡重吸收。缺氧对基因表达的影响是由缺氧诱导因子（HIF）介导的，β 2 AR刺激通过CREB诱导基因表达。它们在离子转运蛋白表达中的作用尚不清楚。由于这两种因子通过共激活蛋白p300和CBP相互作用，因此了解缺氧和p2 AR刺激对肺泡重吸收和离子转运蛋白表达的影响之间的相互作用非常重要，这对于治疗肺水肿具有重要的临床意义。在这个项目中，我们想测试的假设，延长β 2 AR刺激可以防止缺氧诱导的抑制活性和表达的肺泡离子转运蛋白的CREB依赖或非依赖机制。将测试CREB和HIF的作用及其相互作用，以及延长β 2 AR刺激是否阻止缺氧对肺泡转运的影响。实验将在体外对原代大鼠和人肺泡上皮细胞进行，并在用β 2 AR-激动剂特布他林长时间处理后对体内暴露于缺氧的大鼠进行，以具体评价基因表达的作用。HIF和CREB的作用将通过基因沉默来研究。读出参数是离子转运蛋白和β 2AR信号传导的表达和活性。