Table 1.
Primers and probes.
Figure 1.
The effect of incubation of fPCLS on viability, gene expression of fibrosis markers and collagen 1 protein expression.
The effect of incubation up to 48(A) (n = 12), on the gene expression of fibrosis markers Hsp47, αSma and Pcol1A1 (B) (n = 13) and on the collagen 1 protein expression measured by western blot (pictures are from a representative experiment) (C) (n = 10) of fibrotic PCLS from livers of BDL rats. Error bars represent standard error of the mean (SEM). *p<0.05 vs 0 hours. #p<0.05 vs 1 hour.
Figure 2.
Sirius red staining of control fPCLS and fPCLS incubated with antifibrotic drugs.
Sirius red staining on paraffin embedded, formalin fixed fibrotic PCLS directly after slicing (A), incubated for 1 hour (B), 24 hours (C) and 48 hours (D) and incubated for 48 hours with the compounds imatinib (ima) (E), sorafenib (sor) (F), sunitinib (sun) (G), perindopril (per) (H), valproic acid (vpa) (I), rosmarinic acid (ra) (J), tetrandrine (tet) (K), pirfenidone (pir) (L) and colchicine (col) (M).Quantification of sirius red staining (N). n = 3. Error bars represent SEM. *p<0.05 vs 48 hours.
Figure 3.
Hydroxyproline content of control fPCLS and fPCLS incubated with antifibrotic drugs.
Hydroxyproline content in fPCLS incubated up to 48(A) and with addition of 10 µM imatinib (ima), 2 µM sorafenib (sor), 5 µM sunitinib (sun), 100 µM perindopril (per), 1 mM valproic acid (vpa) 270 µM rosmarinic acid (ra), 100 µM tetrandrine (tet), 2.5 mM perindopril (pir) and 200 nM colchicine (col) (B). n = 3. Error bars represent SEM. *p<0.05 vs 0h (A), *p<0.05 vs ctrl 48h (B).
Figure 4.
The effect of PDGF-inhibitors in fPCLS on viability, gene expression of fibrosis markers and collagen 1 protein expression.
The effect of PDGF-inhibitors imatinib (n = 4), sorafenib (n = 4) and sunitinib (n = 4) on the viability (A), on the gene expression of fibrosis markers Hsp47, αSma and Pcol1A1 (B) and on the collagen 1 protein expression (pictures are from a representative experiment) (C) of fibrotic PCLS from livers from BDL rats incubated for 48 hours. Error bars represent SEM. Significant decrease: *p<0.05 vs 48h, #p<0.05 vs 0 hours, +p<0.05 vs 24 hours.
Figure 5.
The effect of TGFβ-inhibitors in fPCLS on viability, gene expression of fibrosis markers and collagen 1 protein expression.
The effect of TGFβ-inhibitors perindopril (n = 5), valproic acid (n = 4), rosmarinic acid (n = 4), tetrandrine (n = 4) and pirfenidone (n = 5) on the viability (A), on the gene expression of fibrosis markers Hsp47, αSma and Pcol1A1 (B) and on the collagen 1 protein expression (pictures are from a representative experiment) (C) of fPCLS from livers from BDL rats incubated for 48 hours. Error bars represent SEM. Significant decrease: *p<0.05 vs 48h, #p<0.05 vs 0 hours, +p<0.05 vs 24 hours.
Figure 6.
The effect of colchcine in fPCLS on viability, gene expression of fibrosis markers and collagen 1 protein expression.
The effect of colchicine (n = 4) on the viability (A), on the gene expression of fibrosis markers Hsp47, αSma and Pcol1A1 (B) and on the collagen 1 protein expression (pictures are from a representative experiment) (C) of fPCLS from livers from BDL rats incubated for 48 hours. Error bars represent SEM. *p<0.05 vs 48h.