The Novel AT2 Receptor Agonist β-Pro7-AngIII Exerts Cardiac and Renal Anti-Fibrotic and Anti-Inflammatory Effects in High Salt-Fed Mice

Figure 1. Representative (a) bright view and (b) polarized view for interstitial fibrosis sections stained with picrosirius red (indicated by red) in left ventricle (LV) obtained from male FVB/N mice fed on a normal salt (NS, 0.5% NaCl) diet or mice fed on a high salt (HS, 5% NaCl) diet in the presence or absence of β-Pro7-AngIII (0.1 mg/kg/day) +/− PD123319 (1 mg/kg/day). Scale bar for all images = 50 μm. (c) Group data of % area of interstitial fibrosis in LV sections. (d) Total tissue collagen measured by hydroxyproline assay in LV tissue from same animal groups in which picrosirius red staining was performed (n = 7–8 per group). All data are expressed as mean ± s.e.m. * p < 0.05, ** p < 0.01 vs. sham; # p < 0.05, ## p < 0.01 vs. HS; & p < 0.05 vs. HS+ β-Pro7-AngIII, determined by One-Way ANOVA with Tukey correction for multiple comparisons.

Figure 2. (a) Representative images for interstitial fibrosis stained with picrosirius red (indicated by red) in kidney sections obtained from male FVB/N mice fed on a normal salt (NS, 0.5% NaCl) diet or mice fed on a high salt (HS, 5% NaCl) diet in the presence or absence of β-Pro7-AngIII (0.1 mg/kg/day) +/- PD123319 (1 mg/kg/day). Scale bar for all images = 50 μm. (b) Group data of % area of interstitial fibrosis in kidney sections. (c) Total tissue collagen measured by hydroxyproline assay in kidney tissue from same animal groups in which picrosirius red staining was performed (n = 7–8 per group). All data are expressed as mean ± s.e.m. *** p < 0.001 vs. sham; # p < 0.05, ### p < 0.001 vs. HS; && p < 0.01 vs. HS+ β-Pro7-AngIII, determined by One-Way ANOVA with Tukey correction for multiple comparisons.

Figure 3. Representative images for (a) TGF-β1 and (b) α-SMA in cardiac sections obtained from male FVB/N mice fed on a normal salt (NS, 0.5% NaCl) diet or mice fed on a high salt (HS, 5% NaCl) diet in the presence or absence of β-Pro7-AngIII (0.1 mg/kg/day) +/− PD123319 (1 mg/kg/day). Scale bar for all images = 50 μm. Group data of % area of (c) TGF-β1 and (d) α-SMA in cardiac sections (n = 7–8 per group). All data are expressed as mean ± s.e.m. ** p < 0.01 vs. sham; # p < 0.05, ## p < 0.01 vs. HS, determined by One-Way ANOVA with Tukey correction for multiple comparisons.

Figure 4. (a) Representative images for TGF-β1 in kidney sections obtained from FVB/N male mice fed on a normal salt (NS, 0.5% NaCl) diet or mice fed on a high salt (HS, 5% NaCl) diet in the presence or absence of β-Pro7-AngIII (0.1 mg/kg/day) +/− PD123319 (1 mg/kg/day). Scale bar for all images = 50 μm. (b) Group data of % area of TGF-β1 in cardiac sections. (c) Plasma TGF-β1 levels (n = 7–8 per group). All data are expressed as mean ± s.e.m. * p < 0.05 vs. sham; # p < 0.05 vs. HS, determined by One-Way ANOVA with Tukey correction for multiple comparisons.

Figure 5. Representative images for (a) p-IκB and (b) MCP-1 in LV sections obtained from male FVB/N mice fed on a normal salt (NS, 0.5% NaCl) diet or mice fed on a high salt (HS, 5% NaCl) diet in the presence or absence of β-Pro7-AngIII (0.1 mg/kg/day) +/− PD123319 (1 mg/kg/day). Scale bar for all images = 50 μm. Group data of % area of (c) p-IκB and (d) MCP-1 in LV sections from indicated groups (n = 4–6 per group). All data are expressed as mean ± s.e.m. * p < 0.05 vs. sham; ## p < 0.01 vs. HS; && p < 0.01 vs. HS+ β-Pro7-AngIII, determined by One-Way ANOVA with Tukey correction for multiple comparisons.

Figure 6. Representative images for (a) p-IκB and (b) F4/80 in kidney sections obtained from male FVB/N mice fed on a normal salt (NS, 0.5% NaCl) diet or mice fed on a high salt (HS, 5% NaCl) diet in the presence or absence of β-Pro7-AngIII (0.1 mg/kg/day) +/− PD123319 (1 mg/kg/day). Scale bar for all images = 50 μm. Group data of % area of (c) p-IκB and (d) F4/80 in kidney sections from indicated groups (n = 6–8 per group). All data are expressed as mean ± s.e.m. ** p < 0.01, *** p < 0.001 vs. sham; ## p < 0.01 vs. HS; & p < 0.05, && p < 0.01 vs. HS+ β-Pro7-AngIII, determined by One-Way ANOVA with Tukey correction for multiple comparisons.