Effects of endothelin-1 on endothelial progenitor cell function
| العنوان: | Effects of endothelin-1 on endothelial progenitor cell function |
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| المؤلفون: | Brian R. Weil, Owen J. MacEneaney, Christian M. Westby, Erich J. Kushner, Jared J. Greiner, Kyle J. Diehl, Christopher A. DeSouza |
| المصدر: | Clinical Chemistry and Laboratory Medicine. 50 |
| بيانات النشر: | Walter de Gruyter GmbH, 2012. |
| سنة النشر: | 2012 |
| مصطلحات موضوعية: | Adult, Vascular Endothelial Growth Factor A, Endothelium, medicine.medical_treatment, Clinical Biochemistry, Apoptosis, Biology, Endothelial progenitor cell, Andrology, Young Adult, chemistry.chemical_compound, Cell Movement, medicine, Humans, Progenitor cell, Endothelin-1, Stem Cells, Growth factor, Biochemistry (medical), General Medicine, Endothelin 1, Vascular endothelial growth factor, Endothelial stem cell, medicine.anatomical_structure, chemistry, Immunology, cardiovascular system, Endothelium, Vascular, Endothelin receptor |
| الوصف: | Background: Circulating endothelial progenitor cells (EPCs) contribute to vascular endothelial repair. Endothelin (ET)-1 is associated with endothelial damage and atherogenesis. The experimental aim of this study was to determine, in vitro, the effects of ET-1 on the ability of EPCs to form colo nies, migrate, release angiogenic growth factors and resist apoptosis. Methods: Peripheral blood samples were collected from 10 healthy adult humans. Cells with phenotypic EPC char-acteristics were isolated and EPC colony-forming capacity (CFU assay), migratory activity (Boyden chamber), release of angiogenic growth factors (enzyme immunoassay) and apoptosis (TUNEL assay) were determined in the absence and presence of ET-1 (100 pmol). Results: EPC colony-forming units (42 ± 12 vs. 39 ± 1), 1migratory capacity (910 ± 146 vs. 936 ± 148 AU) and release of vascular endothelial growth factor (202.8 ± 68.1 vs. 204.8 ± 69.8 pg/mL) and granulocyte-colony stimulating fac-tor (1294.4 ± 378.3 vs. 1 136.1 ± 310.3 pg/mL) were not signifi -cantly affected by ET-1. EPCs treated with ET-1 demonstrated a 20 % increase (p < 0.05) in cellular apoptosis. The proapop-totic effect of ET-1 was abolished with ET receptor blockade as well as with apocynin, a nicotinamide adenine dinucleotide phosphate (NADPH) inhibitor. Conclusions: These results indicate that ET-1 does not affect EPC colony formation, migratory capacity or angiogenic growth factor release, but does increase EPC susceptibil-ity to apoptosis through an NADPH-dependent mechanism. Increased EPC apoptosis may contribute to the proathero-genic effects of ET-1. Keywords: apoptosis; colony-forming units; endothelin-1; endothelial progenitor cells; migration. Endothelin (ET)-1 is the most potent and predominant endothelin isoform in the human cardiovascular system and is intimately involved in the regulation of vascular tone (1) . Additionally, ET-1 has been linked to the etiology of a number of cardiovascular risk factors as well as the development and progression of atherosclerotic vascular disease (2) . Indeed, upregulation of ET-1 and ET-1 receptors have been reported in atherosclerotic plaque. Circulating endothelial progenitor cells (EPCs) are involved in vascular repair processes. EPCs migrate to local sites of ischemia and endothelial damage, secrete potent angiogenic growth factors, and participate in neovascularization (3) . Impaired EPC function has been linked with accelerated atherosclerotic disease progression and adverse cardiovascular outcomes (4, 5) . Interestingly, EPC dysfunction has been observed in conditions associ-ated with enhanced ET-1 system activation (6) ; suggesting a potential link between ET-1 and EPC function. However, whether ET-1 disrupts EPC function is currently unknown. Accordingly, we determined, in vitro, the effects of ET-1 on the ability of EPCs to form colonies, migrate, release angio-genic growth factors and resist apoptosis. Peripheral blood samples were collected from 10 healthy men (age: 21 – 42 years). Putative EPCs were isolated as pre-viously described (7) . Briefl y, mononuclear cells were iso-lated and plated on human fi bronectin (BD Biosciences, San Jose, CA, USA) coated 6-well plates for 48 h at 37 ° C and non-adherent cells were collected. Endothelial-cell lineage was confi rmed by fl uorescent-activated cell sorting (FACS) analysis utilizing endothelial-specifi c antibodies recogniz-ing cell surface expression of vascular endothelial growth factor receptor-2 (VEGFR2), CD34, and CD133, as well as immunofl uorescent staining for VE-cadherin, Dil-ac-LDL uptake, von Willebrand factor, platelet endothelial cell adhe-sion molecule (PECAM-1), and VEGFR2. For each of the assays described below cells were cultured in the absence and presence of: a) ET-1 (100 pM); b) ET-1 (100 pM) + BQ-123/BQ-788 (ET-1 receptor antagonists: 1 μM each); c) ET -1 (100 pM) + apocynin (an NAPH oxidase inhibitor: 25 μ M). All subjects provided written informed consent according to the guidelines of the University of Colorado, Boulder.T reatment effects were determined by analysis of variance. When indicated by a signifi cant main effect, post hoc compar-isons between treatments were performed using the Newman-Kuels methods. All data are expressed as means ± SEM. Statistical signifi cance was set a priori at p < 0.05. The number of colony-forming units (42 ± 12 vs. 39 ± 11 CFU) and migration to VEGF (910 ± 146 vs. 936 ± 148 RFU) were not signifi cantly different in EPCs treated with ET-1 compared with untreated cells (Figure 1 ). There was no effect |
| تدمد: | 1437-4331 1434-6621 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::0e5035f5e9cd41a7890806043833a600Test https://doi.org/10.1515/cclm-2011-0670Test |
| رقم الانضمام: | edsair.doi.dedup.....0e5035f5e9cd41a7890806043833a600 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 14374331 14346621 |
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