A novel role for the mineralocorticoid receptor in glucocorticoid driven vascular calcification

Abstract

Vascular calcification, which is common in the elderly and in patients with atherosclerosis, diabetes and chronic renal disease, increases the risk of cardiovascular morbidity and mortality. It is a complex, active and highly regulated cellular process that resembles physiological bone formation. It has previously been established that pharmacological doses of glucocorticoids facilitate arterial calcification. However, the consequences for vascular calcification of endogenous glucocorticoid elevation have yet to be established. Glucocorticoids (cortisol, corticosterone) are released from the adrenal gland, but can also be generated within cells from 11-keto metabolites of glucocorticoids (cortisone, 11-dehydrocorticosterone [11-DHC]) by the enzyme, 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1). In the current study we hypothesized that endogenous glucocorticoids facilitate vascular smooth muscle cell (VSMC) calcification and investigated the receptor-mediated mechanism underpinning this process.In vitro studies revealed increased phosphate-induced calcification in mouse VSMCs following treatment for 7 days with corticosterone (100 nM; 7.98 fold; P < 0.01), 11-DHC (100 nM; 7.14 fold; P < 0.05) and dexamethasone (10 nM; 7.16 fold; P < 0.05), a synthetic glucocorticoid used as a positive control. Inhibition of 11β-HSD isoenzymes by 10 μM carbenoxolone reduced the calcification induced by 11-DHC (0.37 fold compared to treatment with 11-DHC alone; P < 0.05). The glucocorticoid receptor (GR) antagonist mifepristone (10 μM) had no effect on VSMC calcification in response to corticosterone or 11-DHC. In contrast, the mineralocorticoid receptor (MR) antagonist eplerenone (10 μM) significantly decreased corticosterone- (0.81 fold compared to treatment with corticosterone alone; P < 0.01) and 11-DHC-driven (0.64 fold compared to treatment with 11-DHC alone; P < 0.01) VSMC calcification, suggesting this glucocorticoid effect is MR-driven and not GR-driven. Neither corticosterone nor 11-DHC altered the mRNA levels of the osteogenic markers PiT-1, Osx and Bmp2. However, DAPI staining of pyknotic nuclei and flow cytometry analysis of surface Annexin V expression showed that corticosterone induced apoptosis in VSMCs.This study suggests that in mouse VSMCs, corticosterone acts through the MR to induce pro-calcification effects, and identifies 11β-HSD-inhibition as a novel potential treatment for vascular calcification.

Osteocalcin Regulates Arterial Calcification Via Altered Wnt Signaling and Glucose Metabolism

“…Primary aortic VSMCs were isolated from 5‐week‐old mice as described previously . Mice were euthanized by cervical dislocation.…”

“…Primary aortic VSMCs were isolated from 5-week-old mice as described previously. (3,12,13) Mice were euthanized by cervical dislocation. The aorta was then dissected, the adventitia removed, and the aorta cut open to expose the endothelial layer.…”

A novel role of cellular interactions in vascular calcification

“…Elevated endogenous glucocorticoids were shown to act through mineralocorticoid receptor and facilitate phosphate-induced VSMC calcification [21]. Administration of dexamethasone reduced the expression of MGP, osteopontin and VC-associated factor mRNA on the pericytes, resulting in increased ALP activity and calcium deposition [20].…”

11β-HSD1 inhibition ameliorates diabetes-induced cardiomyocyte hypertrophy and cardiac fibrosis through modulation of EGFR activity

“…11β-HSD1 is a key enzyme that catalyzes the intracellular conversion of the inactive glucocorticoid cortisone to active cortisol (11-dehyrocorticosterone to corticosterone in rodents). Recent studies have suggested that 11β-hydroxysteroid dehydrogenase-1 (11β-HSD1) is an important risk factor for diabetes and diabetic complications [ 4 – 7 ]. Importantly, it is the activity of 11β-HSD1 that determines the intracellular concentration of active glucocorticoids in the tissues and not the plasma glucocorticoid levels [ 8 ].…”