Induction of 11β-hydroxysteroid dehydrogenase type 1 but not -2 in human aortic smooth muscle cells by inflammatory stimuli

doi:10.1016/S0960-0760(01)00041-3

The Journal of Steroid Biochemistry and Molecular Biology

Volume 77, Issues 2–3, May 2001, Pages 117-122

https://doi.org/10.1016/S0960-0760(01)00041-3 Get rights and content

Abstract

The 11β-hydroxysteroid dehydrogenase (11β-HSD) enzymes catalyze the interconversion of active glucocorticoids (GC) with their inert metabolites, thereby regulating the functional activity of GC. While 11β-HSD type 1 (11β-HSD1) activates GC from their 11-keto metabolites, 11β-HSD type 2 (11β-HSD2) inactivates GC. Here we report that both of these enzymes are expressed in human aortic smooth muscle cells (SMC), and that 11β-HSD1 is more abundant and is differentially regulated relative to 11β-HSD2. Stimulation of SMC with IL-1β or TNFα led to a time- and dose-dependent increase of mRNA levels for 11β-HSD1, while 11β-HSD2 mRNA levels decreased. Parallel enzyme activity studies showed increased conversion of ³H-cortisone to ³H-cortisol but not ³H-cortisol to ³H-cortisone, demonstrating 11β-HSD1 in SMC acts primarily as a reductase. A similar increase of 11β-HSD1 mRNA expression was also found in human bronchial SMC upon stimulation, indicating the regulatory effect is not limited to vascular smooth muscle. Additional parallel studies revealed a similar pattern of induction for 11β-HSD1 and monocyte chemoattractant protein-1, a well-defined proinflammatory molecule. These data suggest 11β-HSD1 may play an important role in regulating inflammatory responses in the artery wall and lung.

Introduction

Glucocorticoids (GC, cortisol in man, corticosterone in rodents) regulate a variety of metabolic and homeostatic processes, including carbohydrate, protein, and lipid metabolism and the suppression of immune and inflammatory responses see review [1], [2]. 11β-hydroxysteroid dehydrogenase (11β-HSD) enzymes catalyze the interconversion of active GC (cortisol and corticosterone) with their inert 11-keto metabolites (cortisone and 11-dehydrocorticosterone), thereby regulating these potent and multifunctional hormones via pre-receptor metabolism see review [3]. Two distinct isoenzymes (11β-HSD type 1 and type 2) catalyze this reaction. 11β-HSD type 1 (11β-HSD1) most commonly activates GC from their 11-keto metabolites and is expressed in a wide range of tissues, including the liver, lung, kidney, gonad, pituitary, and adipose tissue [4], [5]. 11β-HSD type 2 (11β-HSD2) converts GC to 11-keto metabolites and is expressed in placenta and mineralocorticoid-responsive tissues including kidney and colon, where it protects the mineralocorticoid receptor (MR) from occupation by the alternative ligand, GC see review [6]. Genetic disruption of the 11β-HSD2 gene in mice and genetic deficiency in man leads to the syndrome of apparent mineralocorticoid excess (SAME), in which GC occupy MR, causing sodium retention, hypokalemia and hypertension [7], [8].

In addition to the potential for GC to act as mineralocorticoids, GC have long been implicated in various forms of human hypertension (see review [9]). While a high sodium intake may worsen the hypertension seen with chronic GC exposure, the primary rise in blood pressure appears to have its roots in altering the contractile responses within the vascular tree. Vascular smooth muscle cells (SMC) express 11β-HSD1 and -2 [10], [11], [12]. Teelucksingh et al. [13] found that administration of 11β-HSD inhibitor, glycyrrhetinic acid, potentiated cortisol-induced vasoconstriction in man, suggesting 11β-HSD may play a role in regulating arterial function and blood pressure in human. Since vascular SMC are frequently exposed to inflammatory stimuli, we asked whether the expression of 11β-HSD in human aortic SMC is affected by inflammatory stimuli.

Section snippets

Cell culture

Human aortic and bronchial smooth muscle cells were purchased from Clonetics (San Diego, CA). Cells were maintained in smooth muscle cell growth medium (basal medium) with 5% FBS, 0.5 ng/ml hEGF, 5 μg/ml insulin, 2 ng/ml hFGF-B and 0.01% GA-1000 in an atmosphere containing 5% CO₂ and 95% air as suggested by the manufactures. Cells at their 5th passage were used in this study. Cultured cells were harvested and washed once with PBS, and then resuspended in the assay medium (basal medium with 0.5%

Induction of 11β-HSD1 but not 11β-HSD-2 mRNA in human aortic SMC upon stimulation

Expression of 11β-HSD mRNA in human aortic SMC was determined using a real time, fluorescence-based, sensitive PCR technique, TaqMan PCR analysis [14]. Consistent with earlier reports [11], human aortic SMC expressed readily detectable levels of 11β-HSD1. TaqMan analysis yielded an average Ct of 25.1±0.6 (n=4) for 11β-HSD1 in unstimulated SMC. By contrast, expression of 11β-HSD2 was much lower, though its mRNA was also detectable. The average Ct for 11β-HSD2 in unstimulated SMC was 34.6±1.0 (n

Discussion

We have demonstrated for the first time that stimulation of human aortic SMC with IL-1β or TNFα enhances the production of 11β-HSD1. While expression of 11β-HSD1 mRNA was greatly enhanced upon stimulation, the expression of 11β-HSD2 was not up-regulated, rather it decreased. An apparent inhibitory activity on 11β-HSD2 mRNA by the stimulation of IL-1β or TNFα may further increase the activity ratio for 11β-HSD1. Though the mRNA level of 11β-HSD2 is substantially lower than that of 11β-HSD1, the

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Induction of 11β-hydroxysteroid dehydrogenase type 1 but not -2 in human aortic smooth muscle cells by inflammatory stimuli

Abstract

Introduction

Section snippets

Cell culture

Induction of 11β-HSD1 but not 11β-HSD-2 mRNA in human aortic SMC upon stimulation

Discussion

Cell

J. Biol. Chem.

Kidney Int.

Lancet

Trends Cardiovasc. Med.

J. Steroid Biochem. Mol. Biol.

Mol. Cell.

The steroid and thyroid hormone receptor superfamily

Science

Function, gene organization and protein structures of 11beta-hydroxysteroid dehydrogenase isoforms

Eur. J. Biochem.

Immunohistochemical localization of type 1 11beta-hydroxysteroid dehydrogenase in human tissues