Fig. 5

Effect of metformin on bleomycin-induced lung fibrosis development in mice. a Body Wight (BW) changes after BLM treatment. BW at day 0 before treatment was designated as 1.0. *p < 0.05. b Photomicrographs of Masson trichrome and Hematoxylin-Eosin staining of mouse lungs at day 21. Upper panels are low magnification view of Masson trichrome staining. Original magnification × 40. Middle panels are High magnification view of Masson trichrome staining. Original magnification × 100. Lower panels are high magnification view of Hematoxylin-Eosin staining. Original magnification × 100. c Shown in the panel is the average (±SEM) soluble collagen measurement from Sircol assay using control (n = 13), BLM-treated (n = 18), and BLM-treated with subsequent metformin injection mouse lungs (n = 15) at day 21. Open bar is control, filled bar is BLM-treated, and horizontal crosshatched bar is BLM-treated with subsequent metformin injection. *p < 0.05. d Immunohistochemical staining of NOX4, p-SMAD3, αSMA in mouse lungs at day 21. Upper panels are high magnification view of NOX4 staining. Original magnification × 200. Middle panels are High magnification view of p-SMAD3 staining. Original magnification × 400. Lower panels are high magnification view of αSMA staining. Original magnification × 200. Bar = 100 μm e Immunohistochemical staining of NOX4 in human lungs. Upper panels are high magnification view of normal lungs. Original magnification × 200. Lower panels are High magnification view of IPF lungs. Original magnification × 400. Bar = 100 μm f WB using anti-NOX4, and anti-β-actin of cell lysates from normal LF (lane 1, 2, 3) and IPF LF (lane 4, 5, 6). Lower panel is the average (±SEM) taken from three patients shown as relative expression. Open bar is normal LF and filled bar is IPF LF. *p < 0.05