Figure 3

Effects of fluvastatin (Flu) on α-SMA protein expression and pro-fibrogenic gene transcripts in conditioned medium (CM) or TGF-β1-treated HSC-T6 cells. (A) HSC-T6 cells incubated in the conditioned medium (CM) from PA-treated PRHs showed significantly increased α-SMA protein expression, while α-SMA protein expression was not up-regulated in HSC-T6 cells treated directly with PA. TGF-β1 (1 ng/mL) was used as a positive control ^*p < 0.05 vs. control. (B) mRNA expressions of TIMP-1 and α-SMA were increased in HSC-T6 cells incubated with CM, whereas there were no significant differences in the expressions of pro-fibrogenic genes when HSC-T6 cells were treated with PA directly. TGF-β1 (1 ng/mL) was used as a positive control. ^**p < 0.01 vs. control. (C) Treatment with Flu significantly reduced CM- and TGF-β1-induced α-SMA protein expression. Besides, α-SMA protein expression was also suppressed in HSC-T6 cells incubated with Flu (5 μM) alone. ^*p < 0.05 vs. control; ^**p < 0.01 vs. control; ^##p < 0.01 vs. CM treatment; &&p < 0.01 vs. TGF-β1 treatment. (D) Flu treatment attenuated CM-induced mRNA expressions of Col1, TGF-β1 and α-SMA, and TGF-β1-induced mRNA expressions of Col1 and α-SMA. ^**p < 0.01 vs. control; ^#p < 0.05 vs. CM treatment; ^##p < 0.01 vs. CM treatment; &p < 0.05 vs. TGF-β1 treatment. (E & F) α-SMA protein and mRNA expressions were significantly decreased in HSC-T6 cells incubated with CM collected from PA-Flu-treated PRHs compared to those incubated with CM collected from PA-treated PRHs. Furthermore, α-SMA protein and mRNA expressions were also reduced in HSC-T6 cells when treated with CM collected from Flu-treated PRHs compared to those incubated with CM without Flu treatment. ^**p < 0.01 vs. control; ^#p < 0.05 vs. CM (PA 200 μM) treatment; &p < 0.05 vs. CM without Flu treatment. n=3 for all experiments.