Department of Biology, University of Padova, I-35121 Padova, Italy.
Bateson Centre, Department of Biomedical Science, University of Sheffield, Sheffield S10 2TN, United Kingdom.
Proc Natl Acad Sci U S A. 2017 Sep 12;114(37):9948-9953. doi: 10.1073/pnas.1705338114. Epub 2017 Aug 29.
Glucocorticoid (GC) and hypoxic transcriptional responses play a central role in tissue homeostasis and regulate the cellular response to stress and inflammation, highlighting the potential for cross-talk between these two signaling pathways. We present results from an unbiased in vivo chemical screen in zebrafish that identifies GCs as activators of hypoxia-inducible factors (HIFs) in the liver. GCs activated consensus hypoxia response element (HRE) reporters in a glucocorticoid receptor (GR)-dependent manner. Importantly, GCs activated HIF transcriptional responses in a zebrafish mutant line harboring a point mutation in the GR DNA-binding domain, suggesting a nontranscriptional route for GR to activate HIF signaling. We noted that GCs increase the transcription of several key regulators of glucose metabolism that contain HREs, suggesting a role for GC/HIF cross-talk in regulating glucose homeostasis. Importantly, we show that GCs stabilize HIF protein in intact human liver tissue and isolated hepatocytes. We find that GCs limit the expression of Von Hippel Lindau protein (pVHL), a negative regulator of HIF, and that treatment with the c-src inhibitor PP2 rescued this effect, suggesting a role for GCs in promoting c-src-mediated proteosomal degradation of pVHL. Our data support a model for GCs to stabilize HIF through activation of c-src and subsequent destabilization of pVHL.
糖皮质激素(GC)和低氧转录反应在组织稳态中起着核心作用,并调节细胞对应激和炎症的反应,这突出了这两种信号通路之间存在潜在的串扰。我们展示了在斑马鱼中进行的一项无偏化学筛选的结果,该筛选鉴定出 GC 是肝脏中缺氧诱导因子(HIFs)的激活剂。GC 以糖皮质激素受体(GR)依赖性的方式激活了公认的缺氧反应元件(HRE)报告基因。重要的是,GC 在携带 GR DNA 结合域点突变的斑马鱼突变系中激活了 HIF 转录反应,这表明 GR 激活 HIF 信号的一种非转录途径。我们注意到,GC 增加了含有 HRE 的几种关键葡萄糖代谢调节剂的转录,这表明 GC/HIF 串扰在调节葡萄糖稳态中发挥作用。重要的是,我们表明 GC 稳定了完整的人肝组织和分离的肝细胞中的 HIF 蛋白。我们发现 GC 限制了 HIF 的负调节因子 Von Hippel Lindau 蛋白(pVHL)的表达,而 c-src 抑制剂 PP2 的治疗挽救了这种作用,这表明 GC 在促进 c-src 介导的 pVHL 蛋白水解降解中起作用。我们的数据支持了 GC 通过激活 c-src 稳定 HIF,随后使 pVHL 不稳定的模型。