Antifibrogenic effects of canrenone, an antialdosteronic drug, on human hepatic stellate cells.
OBJECTIVE
Several lines of evidence indicate that aldosterone antagonists may exert direct antifibrogenic effects. The aim of this study was to evaluate the possible direct antifibrogenic effects of canrenone, the active metabolite of spironolactone, in activated human hepatic stellate cells.


METHODS
The effects of canrenone were assessed on platelet-derived growth factor-induced mitogenic and chemotactic effects and the increased de novo synthesis of different extracellular matrix components induced by transforming growth factor-beta1.


RESULTS
Canrenone dose-dependently reduced platelet-derived growth factor-induced cell proliferation and motility. This effect was not associated with either changes in the phosphorylation of platelet-derived growth factor receptor and phospholipase C gamma or in the activation of the Ras/extracellular signal-regulated kinase pathway, whereas it was accompanied by a dose-dependent inhibition of platelet-derived growth factor-induced phosphatidylinositol 3-kinase activity. In addition, canrenone inhibited the activity of the Na(+)/H(+) exchanger 1 induced by platelet-derived growth factor. The effect of canrenone on Na(+)/H(+) exchanger 1 activity was reproduced by phosphatidylinositol 3-kinase inhibitors, thus supporting an inhibitory action of canrenone on phosphatidylinositol 3-kinase activity. To further address this possibility, the action of canrenone was compared with that of 2 established Na(+)/H(+) exchanger 1 inhibitors: ethylisopropylamiloride and cariporide. Whereas ethylisopropylamiloride was able to inhibit platelet-derived growth factor-induced phosphatidylinositol 3-kinase activity, cariporide was without any effect. Both compounds reproduced the effects of canrenone on platelet-derived growth factor-induced mitogenesis and chemotaxis. Finally, canrenone was able to reduce transforming growth factor-beta1-induced de novo synthesis of procollagen type I/IV and fibronectin and thrombin-induced hepatic stellate cell contraction.


CONCLUSIONS
These results indicate that canrenone may be active as an antifibrogenic drug.
Caligiuri
Alessandra
A
Dipartimento di Medicina Interna, Università di Firenze, Italy.
De Franco
Raffaella M s
RM
Romanelli
Roberto G
RG
Gentilini
Alessandra
A
Meucci
Marta
M
Failli
Paola
P
Mazzetti
Luca
L
Rombouts
Krista
K
Geerts
Albert
A
Vanasia
Massimo
M
Gentilini
Paolo
P
Marra
Fabio
F
Pinzani
Massimo
M
D000450:Aldosterone / Q000378:metabolism; D000584:Amiloride / Q000031:analogs & derivatives* / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D002192:Canrenone / Q000494:pharmacology*; D002455:Cell Division / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D002478:Cells, Cultured; D016326:Extracellular Matrix Proteins / Q000096:biosynthesis; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; D006146:Guanidines / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration / Q000187:drug effects; D007425:Intracellular Membranes / Q000378:metabolism; D008099:Liver / Q000166:cytology / Q000187:drug effects*; D008103:Liver Cirrhosis / Q000517:prevention & control*; D000451:Mineralocorticoid Receptor Antagonists / Q000494:pharmacology*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D010766:Phosphorylation; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D015398:Signal Transduction; D017923:Sodium-Hydrogen Exchangers / Q000037:antagonists & inhibitors; D013450:Sulfones / Q000494:pharmacology; D013917:Thrombin / Q000494:pharmacology
D016326:Extracellular Matrix Proteins; D006146:Guanidines; D000451:Mineralocorticoid Receptor Antagonists; D010982:Platelet-Derived Growth Factor; D017923:Sodium-Hydrogen Exchangers; D013450:Sulfones; D000450:Aldosterone; D002192:Canrenone; D000584:Amiloride; C093373:cariporide; D019869:Phosphatidylinositol 3-Kinases; D011505:Protein-Tyrosine Kinases; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; C496050:PTK2 protein, human; D013917:Thrombin; D002118:Calcium; C039614:ethylisopropylamiloride