Bone morphogenetic protein-7 represses hepatic stellate cell activation and liver fibrosis via regulation of TGF-β/Smad signaling pathway

Figure 3 Bone morphogenetic protein 7 inhibits hepatic stellate cell activation, myofibroblast differentiation, and collagen formation induced by transforming growth factor-beta 1. A: Western blot analysis of expression of p-Smad3 and p-Smad1/5/8. Serum starved (for 20 h) primary mouse HSCs were treated with transforming growth factor-beta 1 (TGF-β1) (2 ng/mL) followed by exogenous BMP7 at the indicated dose or Vehicle for 24 h, and lysed. Equivalent amount of whole cell detergent lysates were blotted with the indicated antibodies. B and C: Densitometry analysis of the effect of exogenous BMP7 on phosphorylation of Smad3 (p-Smad3) and phosphorylation of Smad1/5/8 (p-Smad1/5/8) at the indicated dose. D: Serum starved (for 20 h) primary mouse HSCs were treated with TGF-β1 (2 ng/mL) followed by exogenous BMP7 (100 ng/mL) or Vehicle for 24 h, and lysed. Equivalent amount of whole cell detergent lysates were blotted with the indicated antibodies. E: Densitometry analysis of the effect of exogenous BMP7 on HSCs myofibroblast differentiation and collagen expression evaluated by alpha-smooth muscle actin (α-SMA) and collagen I (Col I) expression (normalized to GAPDH) in panel D. Data are pooled from at least three independent experiments and represented as the mean ± SE. ^bP < 0.01. BMP7: Bone morphogenetic protein 7; p-Smad3: Phosphorylated Smad3; p-Smad1/5/8: Phosphorylated Smad1/5/8; TGF-β1: Transforming growth factor-beta 1; Col I: Collagen I.