title: Precision-cut liver slices as a model for the early onset of liver fibrosis to test antifibrotic drugs.
abstract: Induction of fibrosis during prolonged culture of precision-cut liver slices (PCLS) was reported. In this study, the use of rat PCLS was investigated to further characterize the mechanism of early onset of fibrosis in this model and the effects of antifibrotic compounds. Rat PCLS were incubated for 48h, viability was assessed by ATP and gene expression of PDGF-B and TGF-β1 and the fibrosis markers Hsp47, αSma and Pcol1A1 and collagen1 protein expressions were determined. The effects of the antifibrotic drugs imatinib, sorafenib and sunitinib, PDGF-pathway inhibitors, and perindopril, valproic acid, rosmarinic acid, tetrandrine and pirfenidone, TGFβ-pathway inhibitors, were determined. After 48h of incubation, viability of the PCLS was maintained and gene expression of PDGF-B was increased while TGF-β1 was not changed. Hsp47, αSma and Pcol1A1 gene expressions were significantly elevated in PCLS after 48h, which was further increased by PDGF-BB and TGF-β1. The increased gene expression of fibrosis markers was inhibited by all three PDGF-inhibitors, while TGFβ-inhibitors showed marginal effects. The protein expression of collagen 1 was inhibited by imatinib, perindopril, tetrandrine and pirfenidone. In conclusion, the increased gene expression of PDGF-B and the down-regulation of fibrosis markers by PDGF-pathway inhibitors, together with the absence of elevated TGF-β1 gene expression and the limited effect of the TGFβ-pathway inhibitors, indicated the predominance of the PDGF pathway in the early onset of fibrosis in PCLS. PCLS appear a useful model for research of the early onset of fibrosis and for testing of antifibrotic drugs acting on the PDGF pathway.
authors: Division of Pharmacokinetics, Toxicology and Targeting, Department of Pharmacy, University of Groningen, The Netherlands.
authors: Inge M
authors: IM
authors: Westra
authors: Division of Pharmaceutical Technology and Biopharmacy, Department of Pharmacy, University of Groningen, The Netherlands.
authors: Dorenda
authors: D
authors: Oosterhuis
authors: Division of Pharmacokinetics, Toxicology and Targeting, Department of Pharmacy, University of Groningen, The Netherlands.
authors: Geny M M
authors: GM
authors: Groothuis
authors: Division of Pharmaceutical Technology and Biopharmacy, Department of Pharmacy, University of Groningen, The Netherlands. Electronic address: p.olinga@rug.nl.
authors: Peter
authors: P
authors: Olinga
mesh_terms: D000818:Animals; D000077214:Becaplermin; D001549:Benzamides / Q000494:pharmacology; D044182:Benzylisoquinolines / Q000494:pharmacology; D002934:Cinnamates / Q000494:pharmacology; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D055513:Connective Tissue Growth Factor / Q000235:genetics / Q000378:metabolism; D053630:Depsides / Q000494:pharmacology; D015536:Down-Regulation; D015870:Gene Expression; D050958:HSP47 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D000068877:Imatinib Mesylate; D008099:Liver / Q000187:drug effects* / Q000378:metabolism; D008103:Liver Cirrhosis / Q000188:drug therapy*; D008297:Male; D008954:Models, Biological; D009536:Niacinamide / Q000031:analogs & derivatives / Q000494:pharmacology; D009924:Organ Culture Techniques; D020913:Perindopril / Q000494:pharmacology; D010671:Phenylurea Compounds / Q000494:pharmacology; D010879:Piperazines / Q000494:pharmacology; D020574:Proto-Oncogene Proteins c-sis / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D011728:Pyridones / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D000077157:Sorafenib; D053773:Transforming Growth Factor beta1 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D014635:Valproic Acid / Q000494:pharmacology; D000097123:Rosmarinic Acid
chemical_list: D000077214:Becaplermin; D001549:Benzamides; D044182:Benzylisoquinolines; D002934:Cinnamates; D024042:Collagen Type I; D055513:Connective Tissue Growth Factor; D053630:Depsides; D050958:HSP47 Heat-Shock Proteins; D000068877:Imatinib Mesylate; D009536:Niacinamide; D020913:Perindopril; D010671:Phenylurea Compounds; D010879:Piperazines; D020574:Proto-Oncogene Proteins c-sis; D011728:Pyridones; D011743:Pyrimidines; D000077157:Sorafenib; D053773:Transforming Growth Factor beta1; D014635:Valproic Acid; D000097123:Rosmarinic Acid; C526316:CCN2 protein, rat; C494662:Serpinh1 protein, rat; C009438:tetrandrine; C093844:pirfenidone
keywords: Antifibrotic drugs; Early onset of fibrosis; Ex vivo model; Liver fibrosis; Precision-cut liver slices