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Reciprocal regulation of 11β-hydroxysteroid dehydrogenase 1 and glucocorticoid receptor expression by dexamethasone inhibits human coronary artery smooth muscle cell proliferation in vitro

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Abstract

The actions of glucocorticoids are mediated, in part, by 11β-hydroxysteroid dehydrogenase 1 (11β-HSD1), which amplifies their effects at the pre-receptor level by converting cortisone to cortisol. Glucocorticoids, such as dexamethasone, inhibit vascular smooth muscle cell proliferation; however, the role of 11β-HSD1 in this response remains unknown. Accordingly, we treated human coronary artery smooth muscle cells (HCSMC) with dexamethasone (10−9–10−6 mol/l) and found that after 72 h dexamethasone increased 11β-HSD1 expression (14.16 ± 1.6-fold, P < 0.001) and activity (6.21 ± 1.2-fold, P < 0.001) in a dose- and time-dependent manner, which was dependent upon glucocorticoid receptor (GR) activation and C/EBPβ and C/EBPδ signaling. As glucocorticoids are known to negatively regulate GR expression, we examined the effect of decreasing 11β-HSD1 expression on GR expression. In HCSMC transfected with 11β-HSD1 siRNA, GR expression was increased; this effect was associated with protein kinase A activation and CREB phosphorylation. To examine the role of 11β-HSD1 in HCSMC proliferation, we decreased 11β-HSD1 expression and stimulated cells with platelet-derived growth factor (PDGF) (10 ng/ml). Decreased 11β-HSD1 expression was associated with increased cell proliferation in the absence of PDGF compared to scrambled control-transfected cells (236.10 ± 13.11%, n = 4, P < 0.001) and this effect was augmented by PDGF. Furthermore, the inhibitory effect of dexamethasone on cellular proliferation was abrogated in 11β-HSD1 siRNA-transfected HCSMC. Downregulation of 11β-HSD1 was associated with decreased p27kip1 expression and increased phosphorylated retinoblastoma protein, consistent with a proliferative response. These findings suggest that 11β-HSD1 plays a role in the effects of glucocorticoids on vascular smooth muscle cell phenotype.

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Acknowledgments

The authors would like to thank Professor Vassilis I. Zannis for thoughtful discussions of the data and Ms. Stephanie Tribuna for expert secretarial assistance. This work was supported by NIH grants HL61795, HL81587, HL70819, HL89734 (JL) HL81110, and HL700819 (JAL).

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  1. Cardiovascular Division, Department of Medicine, Brigham and Women’s Hospital and Harvard Medical School, 77 Avenue Louis Pasteur, NRB 0630 K, Boston, MA, 02115, USA

    George Michas, Marcel Liberman, Kristian C. Becker, Diane E. Handy, Joseph Loscalzo & Jane A. Leopold

  2. Graduate Program in Cellular and Genetic Etiology, Diagnosis and Treatment of Human Disease, University of Crete School of Medicine, Crete, Greece

    George Michas

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  1. George Michas
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Correspondence to Jane A. Leopold.

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11010_2010_592_MOESM1_ESM.ppt

Supplementary Fig. 1 Dexamethasone has no effect on 11β-HSD2, mineralocorticoid receptor, or hexose-6-phosphate dehydrogenase expression. Human coronary artery smooth muscle cells (HCSMC) were treated with increasing concentrations of dexamethasone for 72 h and a 11β-HSD2 protein expression was determined by Western blotting (n = 3). b HCSMC were treated with dexamethasone (10−7 mol/l) for up to 72 h and hexose-6-phosphate dehydrogenase (H6PD) expression was determined by Western blotting (n = 3). c HCSMC or Caki-2 cells were treated with dexamethasone (10−7 mol/l) for up to 24 h and mineralocorticoid receptor (MR) expression was examined by Western blotting (n = 3).Representative blots are shown (PPT 144 kb)

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Michas, G., Liberman, M., Becker, K.C. et al. Reciprocal regulation of 11β-hydroxysteroid dehydrogenase 1 and glucocorticoid receptor expression by dexamethasone inhibits human coronary artery smooth muscle cell proliferation in vitro. Mol Cell Biochem 346, 69–79 (2011). https://doi.org/10.1007/s11010-010-0592-5

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