Premium
Dexamethasone alters messenger RNA levels but not synthesis of collagens, fibronectin, or laminin by cultured rat fat‐storing cells
Author(s) -
Niki T,
Schuppan D,
de Bleser P J,
Vrijsen R,
PipeleersMarichal M,
Beyaert R,
Wisse E,
Geerts A
Publication year - 1996
Publication title -
hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.488
H-Index - 361
eISSN - 1527-3350
pISSN - 0270-9139
DOI - 10.1002/hep.510230651
Subject(s) - fibronectin , laminin , messenger rna , dexamethasone , rna , microbiology and biotechnology , chemistry , protein biosynthesis , endocrinology , medicine , biology , extracellular matrix , biochemistry , gene
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 α1(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 α1(III) mRNA level showed a tendency for stimulation. Dexamethasone stimulated the expression of collagen α1 (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.