Stow Lisa R, Gumz Michelle L, Lynch I Jeanette, Greenlee Megan M, Rudin Alicia, Cain Brian D, Wingo Charles S
Department of Physiology and Functional Genomics, University of Florida, Gainesville, Florida 32610, USA.
J Biol Chem. 2009 Oct 30;284(44):30087-96. doi: 10.1074/jbc.M109.030718. Epub 2009 Jul 28.
Aldosterone and endothelin-1 (ET-1) act on collecting duct cells of the kidney and are important regulators of renal sodium transport and cardiovascular physiology. We recently identified the ET-1 gene (edn1) as a novel aldosterone-induced transcript. However, aldosterone action on edn1 has not been characterized at the present time. In this report, we show that aldosterone stimulated edn1 mRNA in acutely isolated rat inner medullary collecting duct cells ex vivo and ET-1 peptide in rat inner medulla in vivo. Aldosterone induction of edn1 mRNA occurred in cortical, outer medullary, and inner medullary collecting duct cells in vitro. Inspection of the edn1 promoter revealed two putative hormone response elements. Levels of heterogeneous nuclear RNA synthesis demonstrated that edn1 mRNA stimulation occurred at the level of transcription. RNA knockdowns corroborated pharmacological studies and demonstrated both mineralocorticoid receptor and glucocorticoid receptor participated in this response. Aldosterone resulted in dose-dependent nuclear translocation and binding of mineralocorticoid receptor and glucocorticoid receptor to the edn1 hormone response elements. Hormone receptors mediated the association of chromatin remodeling complexes, histone modification, and RNA polymerase II at the edn1 promoter. Direct interaction between aldosterone and ET-1 has important implications for renal and cardiovascular function.
醛固酮和内皮素-1(ET-1)作用于肾集合管细胞,是肾钠转运和心血管生理的重要调节因子。我们最近将ET-1基因(edn1)鉴定为一种新的醛固酮诱导转录本。然而,目前醛固酮对edn1的作用尚未明确。在本报告中,我们表明醛固酮在体外急性分离的大鼠肾内髓集合管细胞中刺激edn1 mRNA表达,在体内大鼠肾内髓中刺激ET-1肽表达。醛固酮对edn1 mRNA的诱导在体外的皮质、外髓和内髓集合管细胞中均有发生。对edn1启动子的检查揭示了两个假定的激素反应元件。异质性核RNA合成水平表明,edn1 mRNA的刺激发生在转录水平。RNA敲低证实了药理学研究,并表明盐皮质激素受体和糖皮质激素受体均参与了这一反应。醛固酮导致盐皮质激素受体和糖皮质激素受体向细胞核转位并与edn1激素反应元件结合。激素受体介导了染色质重塑复合物、组蛋白修饰和RNA聚合酶II在edn1启动子处的结合。醛固酮与ET-1之间的直接相互作用对肾和心血管功能具有重要意义。