title: The cardiotonic steroid hormone marinobufagenin induces renal fibrosis: implication of epithelial-to-mesenchymal transition.
abstract: We recently demonstrated that the cardiotonic steroid marinobufagenin (MBG) induced fibrosis in rat hearts through direct stimulation of collagen I secretion by cardiac fibroblasts. This stimulation was also responsible for the cardiac fibrosis seen in experimental renal failure. In this study, the effect of MBG on the development of renal fibrosis in rats was investigated. Four weeks of MBG infusion triggered mild periglomerular and peritubular fibrosis in the cortex and the appearance of fibrotic scars in the corticomedullary junction of the kidney. MBG also significantly increased the protein levels and nuclear localization of the transcription factor Snail in the tubular epithelia. It is known that activation of Snail is associated with epithelial-to-mesenchymal transition (EMT) during renal fibrosis. To examine whether MBG alone can trigger EMT, we used the porcine proximal tubular cell line LLC-PK1. MBG (100 nM) caused LLC-PK1 cells grown to confluence to acquire a fibroblast-like shape and have an invasive motility. The expressions of the mesenchymal proteins collagen I, fibronectin, and vimentin were increased twofold. However, the total level of E-cadherin remained unchanged. These alterations in LLC-PK1 cells in the presence of MBG were accompanied by elevated expression and nuclear translocation of Snail. During the time course of EMT, MBG did not have measurable inhibitory effects on the ion pumping activity of its natural ligand, Na(+)-K(+)-ATPase. Our data suggest that the MBG may be an important factor in inducing EMT and, through this mechanism, elevated levels of MBG in chronic renal failure may play a role in the progressive fibrosis.
authors.affiliation: Division of Nephrology, Dept. of Medicine, Univ. of Toledo College of Medicine, 3000 Arlington Ave., Toledo Ohio, 43614-2598, USA.
authors.forename: Larisa V
authors.initials: LV
authors.lastname: Fedorova
authors.forename: Vanamala
authors.initials: V
authors.lastname: Raju
authors.forename: Nasser
authors.initials: N
authors.lastname: El-Okdi
authors.forename: Amjad
authors.initials: A
authors.lastname: Shidyak
authors.forename: David J
authors.initials: DJ
authors.lastname: Kennedy
authors.forename: Sandeep
authors.initials: S
authors.lastname: Vetteth
authors.forename: David R
authors.initials: DR
authors.lastname: Giovannucci
authors.forename: Alexei Y
authors.initials: AY
authors.lastname: Bagrov
authors.forename: Olga V
authors.initials: OV
authors.lastname: Fedorova
authors.forename: Joseph I
authors.initials: JI
authors.lastname: Shapiro
authors.forename: Deepak
authors.initials: D
authors.lastname: Malhotra
mesh_terms: D000818:Animals; D002018:Bufanolides / Q000008:administration & dosage / Q000633:toxicity*; D002316:Cardiotonic Agents / Q000008:administration & dosage / Q000633:toxicity*; D002465:Cell Movement / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D054338:Cell Transdifferentiation / Q000187:drug effects*; D024042:Collagen Type I / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000633:toxicity; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D005353:Fibronectins / Q000378:metabolism; D005355:Fibrosis; D015918:Infusion Pumps, Implantable; D007668:Kidney / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D007674:Kidney Diseases / Q000139:chemically induced* / Q000378:metabolism / Q000473:pathology; D018374:LLC-PK1 Cells; D008297:Male; D008648:Mesoderm / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D010042:Ouabain / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction / Q000187:drug effects; D000071250:Snail Family Transcription Factors; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors / Q000378:metabolism; D013552:Swine; D013997:Time Factors; D014157:Transcription Factors / Q000378:metabolism; D014746:Vimentin / Q000378:metabolism
chemical_list: D002018:Bufanolides; D002316:Cardiotonic Agents; D024042:Collagen Type I; D004791:Enzyme Inhibitors; D005353:Fibronectins; C000604450:Snai2 protein, rat; D000071250:Snail Family Transcription Factors; D014157:Transcription Factors; D014746:Vimentin; C093896:marinobufagenin; D010042:Ouabain; D000254:Sodium-Potassium-Exchanging ATPase