Regulation of endothelin-converting enzyme-1 (ECE-1) by the calcimimetic R-568

Martínez-Miguel, Patricia; Medrano-Andrés, Diana; Lopes-Martín, Vanessa; Arribas-Gómez, I.; Rodrı́guez-Puyol, Manuel; Rodríguez‐Puyol, Diego; López‐Ongil, Susana

doi:10.1016/j.phrs.2013.07.006

Cited by 8 publications

(5 citation statements)

References 53 publications

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“…Membrane proteins from EA treated for 16 h with 100 nM calcitriol were isolated as described (16), and then 30 g of this homogenate was incubated with human Big ET-1 (10 nM) for 4 h at 37°C in 250 l of a reaction mixture containing 50 mM Tris·HCl buffer pH 7.0 (14). The reaction was stopped by adding 600 l of cold ethanol (Ϫ20°C).…”

Section: Methodsmentioning

confidence: 99%

The active form of vitamin D, calcitriol, induces a complex dual upregulation of endothelin and nitric oxide in cultured endothelial cells

Martínez-Miguel

Valdivielso

Medrano-Andrés³

et al. 2014

American Journal of Physiology-Endocrinology and Metabolism

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Despite the presence of vitamin D receptor (VDR) in endothelial cells, the effect of vitamin D on endothelial function is unknown. An unbalanced production of vasoactive endothelial factors such as nitric oxide (NO) or endothelin-1 (ET-1) results in endothelial dysfunction, which can alter the normal cardiovascular function. Present experiments were devoted to assess the effect of active vitamin D (calcitriol) on the synthesis of endothelial vasoactive factors. The results were that, in cells, calcitriol increased ET-1 and NO productions, which were measured by ELISA and fluorimetric assay, respectively. Calcitriol also increased endothelin-converting enzyme-1 (ECE-1) and endothelial-nitric oxide synthase (eNOS) activities, their mRNA (qPCR), their protein expressions (Western-blot), and their promoter activities (transfection assays). Calcitriol did not change prepro-ET-1 mRNA. The effect was specific to VDR activation because when VDR was silenced by siRNA, the observed effects disappeared. Mechanisms involved in each upregulation differed. ECE-1 upregulation depended on AP-1 activation, whereas eNOS upregulation depended directly on VDR activation. To evaluate the in vivo consequences of acute calcitriol treatment, normal Wistar rats were treated with a single ip injection of 400 ng/kg calcitriol and euthanized 24 h later. Results confirmed those observed in cells, that production and expression of both factors were increased by calcitriol. Besides, calcitriol-treated rats showed a slight rise in mean blood pressure, which decreased when pretreated with FR-901533, an ECE-1 antagonist. We conclude that calcitriol increases the synthesis of both ET-1 and NO in endothelial cells. However, the ET-1 upregulation seems to be biologically more relevant, as animals acutely treated with calcitriol show slight increases in blood pressure.

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Section: Methodsmentioning

confidence: 99%

The active form of vitamin D, calcitriol, induces a complex dual upregulation of endothelin and nitric oxide in cultured endothelial cells

Martínez-Miguel

Valdivielso

Medrano-Andrés³

et al. 2014

American Journal of Physiology-Endocrinology and Metabolism

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“…Cinacalcet alone slightly increased kidney ETA protein, but the net effect on ETB:ETA ratio was beneficial. Previously, cinacalcet and the consequent reduction in PTH were found to down-regulate the activity of endothelin-converting enzyme 1 (ECE-1), which reduced the circulating levels of ET-1 and lowered BP, despite a compensatory increase in the ECE-1 protein [41]. In another study concerning adenine-induced CRI, the moderate reduction of PTH from 441 to 327 pg/mL with cinacalcet attenuated the endothelial-to-mesenchymal transition in rat kidneys, one of the mechanisms of myofibroblast accumulation in renal fibrogenesis [42].…”

Section: Discussionmentioning

confidence: 99%

Unfavorable Reduction in the Ratio of Endothelin B to A Receptors in Experimental 5/6 Nephrectomy and Adenine Models of Chronic Renal Insufficiency

Törmänen

Lakkisto

Eräranta

et al. 2020

IJMS

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Chronic renal insufficiency (CRI) is characterized by increased endothelin 1 (ET-1) synthesis. We studied rat kidney endothelin receptor A (ETA) and receptor B (ETB) expressions after 12 and 27 weeks of 5/6 nephrectomy, and after 12 weeks of 0.3% adenine diet, representing proteinuric and interstitial inflammation models of CRI, respectively. Uric acid and calcium-phosphate metabolism were modulated after 5/6 nephrectomy, while ETA blocker and calcimimetic were given with adenine. Endothelin receptor mRNA levels were measured using RT-qPCR and protein levels using autoradiography (5/6 nephrectomy) or ELISA (adenine model). Both 12 and 27 weeks after 5/6 nephrectomy, kidney cortex ETA protein was increased by ~60% without changes in ETB protein, and the ETB:ETA ratio was reduced. However, the ETB:ETA mRNA ratio did not change. In the adenine model, kidney ETA protein was reduced by ~70%, while ETB protein was suppressed by ~95%, and the ETB:ETA ratio was reduced by ~85%, both at the protein and mRNA levels. The additional interventions did not influence the observed reductions in the ETB:ETA ratio. To conclude, unfavorable reduction in the ETB:ETA protein ratio was observed in two different models of CRI. Therefore, ETA blockade may be beneficial in a range of diseases that cause impaired kidney function.

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“…EA.hy926, a human endothelial cell line (EC), was obtained from the American Type Culture Collection (Manassas, VA, USA), and was used to analyze the effects induced by MCs from each patient at the endothelial level. The EC was grown in Dulbecco's Modified Eagle Media containing 4.5 g/l of glucose supplemented with 10% fetal bovine serum, 100 U/ml penicillin and 100 μg/ml streptomycin in a 5% CO 2 atmosphere (26). The culture media came from Pierce (Rockford, IL, USA).…”

Section: Methodsmentioning

confidence: 99%

“…Membranes were blocked with 5% (w/v) non-fat dry milk in Tris-buffered saline and Tween 20 (20 mM Tris-HCl pH 7.5, 0.9% NaCl, 0.05% Tween 20) at room temperature, and then incubated for 90 min with 10 μg/ml of the monoclonal anti-ECE-1 antibody (mAb AEC32-236 provided by Dr. Kohei Shimada, Biological Research Laboratories, Sankyo, Tokyo, Japan) or with 1:3000 dilution of monoclonal anti-eNOS antibody (BD Biosciences, Madrid, Spain). ECE-1 and eNOS antibodies were usedto evaluate protein content of ECE-1 and eNOS, respectively (26). After washing in Tris-buffered saline and Tween 20, blots were incubated with 200-fold-diluted horseradish peroxidase-conjugated goat anti-mouse IgG (Pierce, Rockford, IL, USA).…”

Section: Methodsmentioning

confidence: 99%

Influence of Polysulphone-Derived Dialysis Membranes on the Interaction of Circulating Mononuclear Cells with the Endothelium

et al. 2014

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Regulation of endothelin-converting enzyme-1 (ECE-1) by the calcimimetic R-568

Cited by 8 publications

References 53 publications

The active form of vitamin D, calcitriol, induces a complex dual upregulation of endothelin and nitric oxide in cultured endothelial cells

The active form of vitamin D, calcitriol, induces a complex dual upregulation of endothelin and nitric oxide in cultured endothelial cells

Unfavorable Reduction in the Ratio of Endothelin B to A Receptors in Experimental 5/6 Nephrectomy and Adenine Models of Chronic Renal Insufficiency

Influence of Polysulphone-Derived Dialysis Membranes on the Interaction of Circulating Mononuclear Cells with the Endothelium

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