title: Aldosterone induces myofibroblast EGF secretion to regulate epithelial colonic permeability.
abstract: In vivo studies show that raised aldosterone (Aldo) during low-Na adaptation regulates the growth of pericryptal myofibroblasts and reduces the permeability of the colonic epithelium. The aim of this study was to reproduce in vitro the in vivo condition of increased Aldo using human CCD-18Co myofibroblasts and T84 colonic epithelial cells to measure myofibroblast and epithelial proliferation and the expression of intercellular junction proteins. Proliferation was quantified by measuring 5-bromo-2'-deoxyuridine incorporation. The myofibroblast expression of EGF, VEGFa, and transforming growth factor-β1 (TGF-β1) was measured by real-time PCR and the expression of junctional complex proteins by Western blot. Aldo stimulated the proliferation of myofibroblasts by 70% (P < 0.05) and increased EGF mRNA expression by 30% (P < 0.05) without affecting VEGFa and TGF-β1. EGF concentration in the incubation medium increased by 30% (P < 0.05) 24 h after Aldo addition, and these effects were prevented by the addition of spironolactone. Myofibroblast proliferation in response to Aldo was mediated by EGF receptor (EGFR) and involved both MAPKK and phosphatidylinositol 3-kinase pathways. When T84 cells were incubated with medium from myofibroblasts stimulated with Aldo (conditioned medium), the expression of β-catenin and claudin IV was increased by 30% (P < 0.05) and proliferation by 40% (P < 0.05). T84 proliferation decreased when α-EGF, or the EGFR antagonist AG1478, was present. Results in vivo indicate that rats fed a low-salt diet showed an increased expression of EGF and EGFR in the colonic mucosa. These results support the view that changes in colonic permeability during low-Na adaptation are mediated by the EGF secreted by myofibroblasts in response to raised Aldo.
authors: Departament de Fisiologia, Facultat de Farmàcia, Universitat de Barcelona, and Institut de Recerca en Nutrició i Seguretat Alimentària, Facultat de Farmàcia, Universitat de Barcelona, Barcelona, Spain.
authors: Lluïsa
authors: L
authors: Miró
authors: Anna
authors: A
authors: Pérez-Bosque
authors: Mònica
authors: M
authors: Maijó
authors: Concepció
authors: C
authors: Amat
authors: Richard J
authors: RJ
authors: Naftalin
authors: Miquel
authors: M
authors: Moretó
mesh_terms: D000222:Adaptation, Physiological; D000450:Aldosterone / Q000494:pharmacology / Q000502:physiology*; D000818:Animals; D002460:Cell Line; D003106:Colon / Q000166:cytology / Q000378:metabolism*; D017077:Culture Media, Conditioned; D004815:Epidermal Growth Factor / Q000235:genetics / Q000378:metabolism*; D004847:Epithelial Cells / Q000378:metabolism; D066246:ErbB Receptors / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism*; D008297:Male; D058628:Myofibroblasts / Q000378:metabolism*; D010539:Permeability; D051381:Rats; D017207:Rats, Sprague-Dawley; D018161:Receptors, Mineralocorticoid / Q000378:metabolism; D015398:Signal Transduction; D017673:Sodium Chloride, Dietary / Q000008:administration & dosage; D053773:Transforming Growth Factor beta1 / Q000235:genetics / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism
chemical_list: D017077:Culture Media, Conditioned; D018161:Receptors, Mineralocorticoid; D017673:Sodium Chloride, Dietary; D053773:Transforming Growth Factor beta1; D042461:Vascular Endothelial Growth Factor A; C467498:vascular endothelial growth factor A, rat; D000450:Aldosterone; D004815:Epidermal Growth Factor; D066246:ErbB Receptors
keywords: EGF; aldosterone; proliferation