Figure 1

In vivo modulation of hepatic macrophages expression profile by 4Mu. (a) Left in vivo experimental model (6 to 8-week-old male C3Hj/He mice; n = 8/group were injected with TAA (200 mg/kg; i.p.) for 4 weeks, 3 times per week, to induce fibrosis. Then, all mice received an intrahepatic inoculation of 1.25 × 10⁵ syngeneic Hepa129 cells (day 0). After tumor implantation, mice received 200 mg/kg 4Mu in drinking water (day 5). On day 9 (n = 4/group) and day 15 (n = 4/group) after 4Mu initiation, mice were sacrificed and liver samples (n = 4/group) were collected. Right The livers were perfused with collagenase and separated in 3 sections (peri-tumoral, tumoral, and non-tumoral tissue). Isolation of non-parenchymal cells from each tissue section was carried out. (b) Representative dot plots of flow cytometry analysis using BD Accuri C6 propietary software version 1.0.264.21 (www.AccuriCytometers.com) of auto fluorescence (upper left panels), F4/80⁺, CD206⁺ and CD86⁺ on non-parenchymal cells. Bar graphs showed the percentage of CD206⁺ F4/80⁺ (M2), and CD86⁺ F4/80⁺ (M1) cells in tumor tissue on day 9 (***p < 0.001) and day 15 (*p < 0.05); 4Mu vs. saline, Mann–Whitney test. Mϕ type1/type2 proportion was calculated as log10 (CD86⁺/CD206⁺). (c) mRNA expression levels of iNOS, Arg1, IL-1β, IL-10, TNF-α, and TGF-β on isolated Mϕ from tumor tissue samples. iNOS /Arg1 ratio **p < 0.01 and *p < 0.05 4Mu vs. Saline (day 9 and day 15 respectively); Mann–Whitney test; TGF-β, IL-1β and TNF-α *p < 0.05; IL-10 ***p < 0.005; 4Mu vs. saline (day 9); TGF-β, IL-1β and TNF-α ****p < 0.001; 4Mu vs. saline (day 15); two-way ANOVA. Data are expressed as the mean ± SEM. The experiment was carried-out 2 times.