Department of Biochemistry and Molecular Biology, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA.
J Biol Chem. 2010 Jul 2;285(27):20570-9. doi: 10.1074/jbc.M110.119495. Epub 2010 May 6.
Leukotrienes, the lipid inflammatory products derived from arachidonic acid, are involved in the pathogenesis of respiratory and cardiovascular diseases and reactive airway disease in sickle cell disease. Placenta growth factor (PlGF), elaborated from erythroid cells, increased the mRNA expression of 5-lipoxygenase and 5-lipoxygenase-activating protein (FLAP) in human pulmonary microvascular endothelial cells. PlGF-induced both promoter activity and mRNA expression of hypoxia-inducible factor-1alpha (HIF-1alpha), which was abrogated by early growth response-1 (EGR-1) small interfering RNA. PlGF showed a temporal reciprocal relationship in the mRNA levels of EGR-1 and NAB2, the latter a repressor of Egr-1. Moreover, Nab2, but not mutant Nab2, significantly reduced promoter activity and mRNA expression of HIF-1alpha and also reduced expression of the HIF-1alpha target gene FLAP. Furthermore, overexpression of Egr-1 led to increased promoter activities for both HIF-1alpha and FLAP in the absence of PlGF. Additionally, the Egr-1-mediated induction of HIF-1alpha and FLAP promoters was reduced to basal levels by EGR-1 small interfering RNA. The binding of Egr-1 to HIF-1alpha promoter was corroborated by electrophoretic mobility shift assay and chromatin immunoprecipitation assay, which showed increased Egr-1 binding to the HIF-1alpha promoter in response to PlGF stimulation. These studies provide a novel mechanism for PlGF-mediated regulation of HIF-1alpha via Egr-1, which results in increased FLAP expression. This study provides a new therapeutic target, namely Egr-1, for attenuation of elevated leukotriene levels in patients with sickle cell disease and other inflammatory diseases.
白三烯是花生四烯酸衍生的脂质炎症产物,参与呼吸和心血管疾病以及镰状细胞病中的反应性气道疾病的发病机制。胎盘生长因子(PlGF)由红系细胞产生,可增加人肺微血管内皮细胞中 5-脂氧合酶和 5-脂氧合酶激活蛋白(FLAP)的 mRNA 表达。PlGF 诱导低氧诱导因子-1α(HIF-1α)的启动子活性和 mRNA 表达,而早期生长反应-1(EGR-1)的小干扰 RNA 可阻断这一作用。PlGF 在 EGR-1 和 NAB2 的 mRNA 水平上表现出时间上的相互关系,后者是 Egr-1 的抑制剂。此外,Nab2 而非突变 Nab2 显著降低 HIF-1α的启动子活性和 mRNA 表达,也降低 HIF-1α 靶基因 FLAP 的表达。此外,在没有 PlGF 的情况下,过表达 Egr-1 导致 HIF-1α 和 FLAP 的启动子活性增加。此外,通过 EGR-1 小干扰 RNA 将 Egr-1 介导的 HIF-1α 和 FLAP 启动子的诱导降低至基础水平。电泳迁移率变动分析和染色质免疫沉淀分析证实了 Egr-1 与 HIF-1α 启动子的结合,该结合在 PlGF 刺激下增加了 Egr-1 与 HIF-1α 启动子的结合。这些研究提供了一种新的机制,即 PlGF 通过 Egr-1 介导 HIF-1α 的调节,导致 FLAP 表达增加。该研究为镰状细胞病和其他炎症性疾病患者中升高的白三烯水平的衰减提供了一个新的治疗靶点,即 Egr-1。