Fig 1.
H&E-stained paraffin-embedded rat bladder sections.
In PBOO rats, significant detrusor muscle hypertrophy, alongside a progressive increase in fibrosis (dyed pale pink, blue arrow) and loss of normal muscle tissue (dyed purple or red, black arrow)architecture was observable at 4 and 8 weeks of PBOO. Fibrosis was inhibited in the STS group at (a) week 4 in Sham group, (b) week 4 after PBOO, (c) week 4 after PBOO+STS treatment, (d) week 8 after Sham group, (e) week 8 after PBOO, and(f) week 8 after PBOO+STS treatment. The PBOO and STS groups show detrusor hypertrophy and collagen deposition. Original magnifications, ×40.
Fig 2.
Masson’s trichrome-stained sections show that PBOO induces bladder fibrosis and STS suppresses bladder fibrosis.
Collagen fibril is represented by the green staining and muscle is purple (A) Representative Masson’s trichrome–stained sections. (a) Sham group at 4 weeks, (b) week 4 after PBOO, (c) week 4 after PBOO+STS treatment, (d) week 8 after Sham group, (e) 8 weeks after PBOO, (f) 8 weeks after PBOO+STS treatment. (B) Fibrotic area scores of the bladder tissues. Each bar represents the mean±SD of eight rats. *P<0.05 vs. Sham group (n = 8); #P<0.05 vs PBOO group. Original magnifications, ×400.
Fig 3.
RT- PCR of a-SMA (Acta2), collagen I (Col1a1), collagen III (Col3a1) mRNA expression in vivo after STS treatment in the PBOO model.
(a) Acta2 mRNA, (b) Col1a1 mRNA, (c) Col3a1 mRNA. Acta2, Col1a1 and Col3a1 mRNA expression was significantly higher in PBOO rats at weeks 4 and 8 compared with Sham rats (P<0.05, n = 8). STS treatment led to decreased Acta2, Col1a1, Col3a1 mRNA expression (P<0.05, n = 8). Each bar represents the mean±SD for eight rats. *P<0.05 vs. sham group (n = 8); #P<0.05 vs. PBOO group.
Fig 4.
Western blotting analyses of a-SMA, collagen I, and collagen III protein expression in vivo after STS treatment in the PBOO model.
a-SMA, collagen I and collagen III expression was significantly higher in PBOO rats at weeks 4 and 8 compared with the Sham group (P<0.05, n = 8). Each bar represents the mean±SD of 8 rats. *P<0.05 vs. sham group (n = 8); #P<0.05 vs. PBOO group.
Fig 5.
STS treatment suppresses a-SMA expression in the PBOO model.
(A) Representative a-SMA-immunostained rat bladder sections. (a) Sham group at 4 weeks, (b) week 4 after PBOO, (c) week 4 after PBOO+STS treatment, (d) week 8 after Sham group, (e) week 8 after PBOO, (f) week 8 after PBOO+STS treatment. a-SMA expression was significantly higher in PBOO rats at weeks 4 and 8 compared with Sham groups (P<0.05, n = 8). STS treatment induced decreased a- SMA expression (P<0.05, n = 8). (B) Densitometric analyses performed from 8 independent experiments. Each bar represents the mean±SD for 8 rats. *P<0.05 vs. sham group (n = 8); #P<0.05 vs. PBOO group. Original magnifications, ×400. IOD, integrated optical density.
Fig 6.
STS treatment suppresses collagen I expression in the PBOO model.
(A) Representative collagen I-immunostained rat bladder sections. (a) Sham group at 4 weeks, (b) week 4 after PBOO, (c) week 4 after PBOO+STS treatment, (d) week 8 after Sham group, (e) week 8 after PBOO, (f) week 8 after PBOO+STS treatment. Collagen I expression was significantly higher in PBOO rats at weeks 4 and 8 compared with the Sham groups (P<0.05, n = 8). STS treatment induced decreased collagen I expression (P<0.05, n = 8). (B) Densitometric analyses were performed from 8 independent experiments. Each bar represents the mean±SD of 8 rats. *P<0.05 vs. Sham group (n = 8); #P<0.05 vs. PBOO group. Original magnifications, ×400.
Fig 7.
STS treatment suppresses collagen III expression in the PBOO model.
(A) Representative collagen III immunostained rat bladder sections. (a) Sham group at 4 weeks, (b) week 4 after PBOO, (c) week 4 after PBOO+STS treatment, (d) week 8 after Sham group, (e) week 8 after PBOO, (f) week 8 after PBOO+STS treatment. Collagen III expression was significantly higher in the PBOO rats at weeks 4 and 8 compared with the Sham groups (P<0.05, n = 8). STS treatment induced decreased collagen III (P<0.05, n = 8). (B) Densitometric analyses performed from 8 independent experiments. Each bar represents the mean±SD of 8 rats. *P<0.05 vs. Sham group (n = 8); #P<0.05 vs. PBOO group. Original magnifications, ×400.
Fig 8.
RT-PCR of (a)Tgfb1, (b)Smad2 and (c)Smad3 mRNA expression in vivo after STS treatment in PBOO model.
Compared with the Sham group, Tgfb1, Smad2 and Smad3 mRNA expression in the PBOO group was not significantly different. Densitometric analyses were performed from 8 independent experiments. Each bar represents the mean±S.D of 8 rats. *P<0.05 vs. sham group (n = 8); #P<0.05 vs. PBOO group.
Fig 9.
Western blotting analyses of TGF-β1, Smad2 and Smad3 protein expression in vivo after STS treatment in the PBOO model.
Compared to the Sham group, TGF-β1 and phosphorylated Smad2/3 expression but not total Smad2/3 expression in PBOO was significantly higher at weeks 4 and 8 (P<0.05, n = 8). Densitometric analyses performed from 8 independent experiments. Each bar represents the mean±SD of 8 rats. *P<0.05 vs. sham group (n = 8); #P<0.05 vs. PBOO group.
Fig 10.
STS treatment suppresses TGF-β1 expression in the PBOO model.
(A) Representative TGF-β1-immunostained rat bladder sections. (a) Sham group at 4 weeks, (b) week 4 after PBOO, (c) week 4 after PBOO+STS treatment, (d) week 8 after Sham group, (e) week 8 after PBOO, (f) week 8 after PBOO+STS treatment. TGF-β1 expression was significantly higher in the PBOO rats at weeks 4 and 8 compared with the Sham groups (P<0.05, n = 8). STS Treatment induced decreased TGF-β1 expression (P<0.05, n = 8). (B) Densitometric analyses performed from 8 independent experiments. Each bar represents the mean±SD for 8 rats. *P<0.05 vs. sham group (n = 8); #P<0.05 vs. PBOO group. Original magnifications, ×400.
Fig 11.
STS treatment suppresses expression of phosphorylated Smad2 in the PBOO model.
(A) Representative Phosphorylated Smad2-immunostained rat bladder sections. (a) Sham group at 4 weeks, (b) week 4 after PBOO, (c) week 4 after PBOO+STS treatment, (d) week 8 after Sham group, (e) week 8 after PBOO, (f) week 8 after PBOO+STS treatment. Expression of Phosphorylated Smad2 was significantly higher in the PBOO rats at weeks 4 and 8 compared with the Sham groups (P<0.05, n = 8). STS Treatment induced decreased Phosphorylated Smad2 (P<0.05, n = 8). (B) Densitometric analyses performed from 8 independent experiments. Each bar represents the mean±SD for 8 rats. *P<0.05 vs. sham group (n = 8); #P<0.05 vs. PBOO group. Original magnifications, ×400. IOD, integrated optical density.
Fig 12.
STS treatment suppresses expression of phosphorylated Smad3 in the PBOO model.
(A) Representative phosphorylated Smad3-immunostained rat bladder sections. (a) Sham group at 4 weeks, (b) week 4 after PBOO, (c) week 4 after PBOO+STS treatment, (d) week 8 after Sham group, (e) week 8 after PBOO, (f) week 8 after PBOO+STS treatment. Expression of phosphorylated Smad3 was significantly higher in PBOO rats at weeks 4 and 8 compared with the Sham group (P<0.05, n = 8). STS treatment induced decreased expression of phosphorylated Smad3 (P<0.05, n = 8). (B) Densitometric analyses performed from 8 independent experiments. Each bar represents the mean±SD of 8 rats. *P<0.05 vs. sham group (n = 8); #P<0.05 vs. PBOO group. Original magnifications, ×400. IOD, integrated optical density.