Influence of aldosterone on collagen synthesis and proliferation of rat cardiac fibroblasts.
1. Previous in vivo studies in men and experimental animal models have shown that hyperaldosteronemia is correlated with cardiac fibrosis due to increased total collagen synthesis. As yet, it is unclear whether aldosterone has direct pro-fibrogenic effect on cardiac fibroblasts, the fibrogenic effector cell in the myocardium, and if so which procollagens specifically are synthesized at higher rates. 2. The present study aims at establishing whether de novo collagen synthesis by cardiac fibroblasts is enhanced following exposure for 2x24 h to pharmacological (10(-7) - 10(-8) M), near-physiological (10(-9) M) or physiological (10(-10) - 10(-11) M) aldosterone concentrations. During the last 24 h, cells were metabolically labelled with [35S]-methionine/[35S]-cysteine. Labelled procollagens were immunoprecipitated quantitatively using antibodies against specific procollagens. Contrary to expectations, 10(-7) M aldosterone inhibited significantly de novo synthesis of procollagens type I and IV (-35% and -42%, respectively). For procollagen type III, only a tendency towards inhibition was observed. At lower concentrations of aldosterone (10(-8) - 10(-10) M), synthesis of procollagens type I, III or IV was unaffected. 3. Cellular DNA synthesis under influence of aldosterone was evaluated by measuring BrdU incorporation. Cells were treated with aldosterone, while BrdU was added during the last 16 h of treatment. Aldosterone had no demonstrable effect on cellular proliferation. 4. Reverse transcription-polymerase chain reaction (RT - PCR) clearly demonstrated the presence of mineralocorticoid receptor mRNA in cardiac fibroblasts. 5. In spite of the expression of the mineralocorticoid receptor by cultured cardiac fibroblasts, the pro-fibrogenic effect of aldosterone as observed in vivo, is not likely to be due to a direct effect of this hormone in cardiac fibroblasts.
Rombouts
K
K
Laboratory for Molecular Liver Cell Biology, Faculty of Medicine and Pharmacy, Free University of Brussels (VUB), Belgium. krom@cyto.vub.ac.be
Wielant
A
A
Hellemans
K
K
Schuppan
D
D
Geerts
A
A
D000199:Actins / Q000032:analysis; D000450:Aldosterone / Q000494:pharmacology*; D000818:Animals; D002455:Cell Division / Q000187:drug effects*; D003094:Collagen / Q000096:biosynthesis / Q000187:drug effects*; D004247:DNA / Q000096:biosynthesis / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D015870:Gene Expression; D007150:Immunohistochemistry; D008297:Male; D009130:Muscle, Smooth / Q000737:chemistry; D009206:Myocardium / Q000737:chemistry / Q000166:cytology; D011233:Precipitin Tests; D011347:Procollagen / Q000096:biosynthesis / Q000187:drug effects; D014176:Protein Biosynthesis; D011506:Proteins / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D018161:Receptors, Mineralocorticoid / Q000235:genetics
D000199:Actins; D011347:Procollagen; D011506:Proteins; D012333:RNA, Messenger; D018161:Receptors, Mineralocorticoid; D000450:Aldosterone; D003094:Collagen; D004247:DNA