title: Dexamethasone alters messenger RNA levels but not synthesis of collagens, fibronectin, or laminin by cultured rat fat-storing cells.
abstract: Glucocorticoids have been shown to suppress collagen synthesis and gene expression by fibroblasts. However, little is known about their effects on fat-storing cells, the major matrix-producing cells in liver fibrosis. In this study we investigated the effect of dexamethasone on the extracellular matrix expression by cultured rat fat-storing cells. Fat-storing cells were isolated from male Wistar rats by collagenase/pronase digestion and purified by density gradient centrifugation. Fat-storing cells in early primary culture (3-day-old, representing a relatively quiescent phenotype) and in subculture (one passage, about 2-week-old, representing an activated phenotype) were treated with 10(-6) mol/L dexamethasone for messenger RNA (mRNA) study or with 10(-8) to 10(-6) mol/L dexamethasone for protein study. Expression of collagen type I, III, IV, fibronectin, and laminin was analyzed at the mRNA level by Northern hybridization, and at the protein level by metabolic labeling and immunoprecipitation. Dexamethasone had a variable effect on the expression of collagen alpha1(I) mRNA level. While a tendency for modest suppression was observed (5%-50%) in primary cells, the difference was not statistically significant. Variable response was observed in subcultured cells. Collagen alpha1(III) mRNA level showed a tendency for stimulation. Dexamethasone stimulated the expression of collagen alpha1 (IV), fibronectin, and laminin B1 mRNA levels by 1.4-, 2.4-, and 1.6-fold respectively, in primary fat-storing cells. Subcultured cells showed a similar response, but the magnitude of stimulation was more variable than that of primary cells. Unexpectedly, at the protein level dexamethasone had no effect on the expression of these proteins. Our results indicate that glucocorticoids do not possess a net suppressive effect on extracellular matrix synthesis by fat-storing cells. Beneficial effects of glucocorticoids may be attributable to other mechanisms of action, such as their anti-inflammatory effect.
authors.affiliation: Laboratory for Cell Biology and Histology,Faculty of Medicine and Pharmacy, Free University of Brussels, Belgium.
authors.forename: T
authors.initials: T
authors.lastname: Niki
authors.forename: D
authors.initials: D
authors.lastname: Schuppan
authors.forename: P J
authors.initials: PJ
authors.lastname: de Bleser
authors.forename: R
authors.initials: R
authors.lastname: Vrijsen
authors.forename: M
authors.initials: M
authors.lastname: Pipeleers-Marichal
authors.forename: R
authors.initials: R
authors.lastname: Beyaert
authors.forename: E
authors.initials: E
authors.lastname: Wisse
authors.forename: A
authors.initials: A
authors.lastname: Geerts
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D003094:Collagen / Q000096:biosynthesis* / Q000235:genetics*; D003907:Dexamethasone / Q000494:pharmacology*; D005353:Fibronectins / Q000096:biosynthesis* / Q000235:genetics*; D015870:Gene Expression / Q000187:drug effects; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000096:biosynthesis / Q000235:genetics; D007797:Laminin / Q000096:biosynthesis* / Q000235:genetics*; D050356:Lipid Metabolism; D008099:Liver / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D008297:Male; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism*; D051381:Rats
chemical_list: D005353:Fibronectins; D007797:Laminin; D012333:RNA, Messenger; D003907:Dexamethasone; D003094:Collagen; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases