Aguado A, Rodríguez C, Martínez-Revelles S, Avendaño M S, Zhenyukh O, Orriols M, Martínez-González J, Alonso M J, Briones A M, Dixon D A, Salaices M
Departamento de Farmacología, Universidad Autónoma de Madrid, Instituto de Investigación Hospital La Paz (IdiPAZ), Madrid, Spain.
Centro de Investigación Cardiovascular (CSIC-ICCC), IIB-Sant Pau, Barcelona, Spain.
Br J Pharmacol. 2015 Jun;172(12):3028-42. doi: 10.1111/bph.13103. Epub 2015 Mar 27.
Angiotensin II (AngII) and IL-1β are involved in cardiovascular diseases through the induction of inflammatory pathways. HuR is an adenylate- and uridylate-rich element (ARE)-binding protein involved in the mRNA stabilization of many genes. This study investigated the contribution of HuR to the increased expression of COX-2 induced by AngII and IL-1β and its consequences on VSMC migration and remodelling.
Rat and human VSMCs were stimulated with AngII (0.1 μM) and/or IL-1β (10 ng · mL(-1)). Mice were infused with AngII or subjected to carotid artery ligation. mRNA and protein levels were assayed by quantitative PCR, Western blot, immunohistochemistry and immunofluorescence. Cell migration was measured by wound healing and transwell assays.
In VSMCs, AngII potentiated COX-2 and tenascin-C expressions and cell migration induced by IL-1β. This effect of AngII on IL-1β-induced COX-2 expression was accompanied by increased COX-2 3' untranslated region reporter activity and mRNA stability, mediated through cytoplasmic HuR translocation and COX-2 mRNA binding. These effects were blocked by ERK1/2 and HuR inhibitors. VSMC migration was reduced by blockade of ERK1/2, HuR, COX-2, TXAS, TP and EP receptors. HuR, COX-2, mPGES-1 and TXAS expressions were increased in AngII-infused mouse aortas and in carotid-ligated arteries. AngII-induced tenascin-C expression and vascular remodelling were abolished by celecoxib and by mPGES-1 deletion.
The synergistic induction of COX-2 by AngII and IL-1β in VSMCs involves HuR through an ERK1/2-dependent mechanism. The HuR/COX-2 axis participates in cell migration and vascular damage. HuR might be a novel target to modulate vascular remodelling.
血管紧张素II（AngII）和白细胞介素-1β（IL-1β）通过诱导炎症途径参与心血管疾病。HuR是一种富含腺苷酸和尿苷酸元件（ARE）的结合蛋白，参与许多基因的mRNA稳定。本研究调查了HuR对AngII和IL-1β诱导的COX-2表达增加的作用及其对血管平滑肌细胞（VSMC）迁移和重塑的影响。
用AngII（0.1μM）和/或IL-1β（10 ng·mL⁻¹）刺激大鼠和人VSMC。给小鼠输注AngII或进行颈动脉结扎。通过定量PCR、蛋白质印迹、免疫组织化学和免疫荧光检测mRNA和蛋白质水平。通过伤口愈合和Transwell实验测量细胞迁移。
在VSMC中，AngII增强了IL-1β诱导的COX-2和肌腱蛋白-C表达以及细胞迁移。AngII对IL-由1β诱导的COX-2表达的这种作用伴随着COX-2 3'非翻译区报告基因活性和mRNA稳定性的增加，这是通过细胞质HuR易位和COX-2 mRNA结合介导的。这些作用被ERK1/2和HuR抑制剂阻断。阻断ERK1/2、HuR、COX-2、TXAS、TP和EP受体可减少VSMC迁移。在输注AngII的小鼠主动脉和颈动脉结扎动脉中，HuR、COX-2、mPGES-1和TXAS表达增加。塞来昔布和mPGES-1缺失消除了AngII诱导的肌腱蛋白-C表达和血管重塑。
AngII和IL-1β在VSMC中协同诱导COX-2涉及通过ERK1/2依赖性机制的HuR。HuR/COX-2轴参与细胞迁移和血管损伤。HuR可能是调节血管重塑的新靶点。
