Lister Kerrie, Autelitano Dominic J, Jenkins Anna, Hannan Ross D, Sheppard Karen E
Baker Heart Research Institute, Prahran, Victoria, Australia.
Cardiovasc Res. 2006 Jun 1;70(3):555-65. doi: 10.1016/j.cardiores.2006.02.010. Epub 2006 Feb 14.
Mineralocorticoids and glucocorticoids have been implicated in the pathogenesis of cardiac diseases; however, both in vivo and in vitro studies indicate that changes in the cellular milieu of either the cardiomyocyte and/or cells of the vasculature is required for corticosteroid signalling to be pathological. The aim of the current study was to directly address whether signalling pathways that are activated during myocyte hypertrophy alter corticosteroid signalling and thus enable these steroids to significantly impact on the hypertrophic response.
Neonatal rat ventricular cardiomyocytes were treated with phenylephrine or phorbol ester for 48 h to induce myocyte hypertrophy. Following treatment, the expression of glucocorticoid receptor, mineralocorticoid receptor, and 11beta-hydroxysteroid dehydrogenase were determined by ribonuclease protection assay. In addition, the activity of 11beta-hydroxysteroid dehydrogenase and the ability of glucocorticoid and mineralocorticoid receptors to induce serum- and glucocorticoid-induced kinase 1 (SGK1) gene transcription were assessed. Corticosteroid effects on phenylephrine and phorbol ester-induced hypertrophy were determined by measuring atrial natriuretic peptide (ANP) mRNA expression, protein synthesis, or induction of rDNA transcription.
Incubation of cardiomyocytes with phenylephrine and phorbol ester for 48 h led to a hypertrophic response with an associated 8- to 12-fold increase in ANP mRNA and 2-fold increase in rDNA transcription. Cardiomyocyte hypertrophy led to a significant 2-fold increase in glucocorticoid receptor and mineralocorticoid receptor expression that resulted in enhanced receptor signaling as judged via the ability of corticosterone and aldosterone to induce SGK1 gene transcription. 11beta-Hydroxysteroid dehydrogenase2 was not detected in normal or hypertrophied cardiomyocytes, and 11beta-hydroxysteroid dehydrogenase exclusively demonstrated reductase activity, converting the inactive 11-ketometabolite back to active glucocorticoid. 11beta-Hydroxysteroid dehydrogenase1 expression and reductase activity were increased with phorbol ester-induced hypertrophy but not phenylephrine-induced hypertrophy. In basal cardiomyocytes, either aldosterone or corticosterone induced only a minor increase in ANP mRNA and protein synthesis; however, in cardiomyocytes primed with phenylephrine, both corticosteroids significantly potentiated phenylephrine-mediated effects via activation of the glucocorticoid receptor.
In the present study we demonstrate that significant cross talk exists in the cardiomyocyte between corticosteroid receptor-activated pathways and both protein kinase C and alpha-adrenergic signalling. Cellular changes associated with the hypertrophic response promote corticosteroid signalling and allow for corticosteroid-mediated potentiation of alpha-adrenergic receptor signalling. Such augmentation of cardiomyocyte hypertrophy may in part explain the role that corticosteroid hormones play in the pathophysiological progression of heart disease.
盐皮质激素和糖皮质激素与心脏疾病的发病机制有关;然而,体内和体外研究均表明,皮质类固醇信号转导要产生病理作用,心肌细胞和/或脉管系统细胞的细胞环境变化是必需的。本研究的目的是直接探讨在心肌细胞肥大过程中激活的信号通路是否会改变皮质类固醇信号转导,从而使这些类固醇能够显著影响肥大反应。
用去甲肾上腺素或佛波酯处理新生大鼠心室心肌细胞48小时以诱导心肌细胞肥大。处理后,通过核糖核酸酶保护试验测定糖皮质激素受体、盐皮质激素受体和11β-羟基类固醇脱氢酶的表达。此外,评估11β-羟基类固醇脱氢酶的活性以及糖皮质激素和盐皮质激素受体诱导血清和糖皮质激素诱导激酶1(SGK1)基因转录的能力。通过测量心房利钠肽(ANP)mRNA表达、蛋白质合成或rDNA转录的诱导来确定皮质类固醇对去甲肾上腺素和佛波酯诱导的肥大的影响。
用去甲肾上腺素和佛波酯孵育心肌细胞48小时导致肥大反应,ANP mRNA增加8至12倍,rDNA转录增加2倍。心肌细胞肥大导致糖皮质激素受体和盐皮质激素受体表达显著增加2倍,这导致通过皮质酮和醛固酮诱导SGK1基因转录的能力判断受体信号增强。在正常或肥大的心肌细胞中未检测到11β-羟基类固醇脱氢酶2,且11β-羟基类固醇脱氢酶仅表现出还原酶活性,将无活性的11-酮代谢物转化回活性糖皮质激素。佛波酯诱导的肥大增加了11β-羟基类固醇脱氢酶1的表达和还原酶活性,但去甲肾上腺素诱导的肥大未增加。在基础心肌细胞中,醛固酮或皮质酮仅诱导ANP mRNA和蛋白质合成轻微增加;然而,在用去甲肾上腺素预处理的心肌细胞中,两种皮质类固醇均通过激活糖皮质激素受体显著增强去甲肾上腺素介导的作用。
在本研究中,我们证明心肌细胞中皮质类固醇受体激活途径与蛋白激酶C和α-肾上腺素能信号转导之间存在显著的相互作用。与肥大反应相关的细胞变化促进皮质类固醇信号转导,并允许皮质类固醇介导的α-肾上腺素能受体信号转导增强。心肌细胞肥大的这种增强可能部分解释了皮质类固醇激素在心脏病病理生理进展中所起的作用。