Figure 7. Liver fibrosis is decreased in NOX2^-/- mice.

(A) Wt or NOX2^-/- mice were bile duct ligated (BDL) then sacrificed after 3 weeks (in the case of some NOX2^-/- mice after 6 weeks) following surgery. Picrosirius staining was done to assess fibrosis stage. In NOX2^-/- animals the fibrosis stage was significantly lower compared to that of wt animals following BDL (Figure 7A b, c, d). NOX2^-/- mice survived longer with less fibrosis compared to wt animals (Figure 7A, b, d). ALT and bilirubin values were increased in both the wt and NOX2^-/- BDL animals (*p<0.05). (B) Mice undergone BDL were injected with GdCl₃, throughout the experiment, to inhibit macrophage function. The fibrosis decreased in GdCl₃-injected wt animals after BDL (Figure 7B, a) compared to PBS-injected controls (Figure 7A, b), while no significant change was seen in NOX2^-/- livers, bars=50μm. Real-time PCR was performed using collagen IA1 specific primers, and OH-proline incorporation assay to assess the amount of collagen in all samples. In NOX2^-/- BDL mice the expression of collagen IA1 (**p≤0.01), and OH-proline incorporation significantly decreased (*p<0.05) compared to wt BDL animals. Inhibiting macrophages decreased collagen IA1 expression (*p<0.05) and OH-proline incorporation (*p<0.05) in wt BDL animals, compared to PBS-injected animals, but not to the extent seen in NOX2^-/- BDL mice. In GdCl₃-injected mice the ALT values were overall lower (*p<0.05) than in PBS-injected mice however, the values among GdCl₃-injected wt and NOX2^-/- animals were not significantly different.