Effect of Ang-(1–7) treatment on apoptosis, fibrosis and inflammatory processes in wild-type mice with unilateral ureteral obstruction (UUO).
Wild-type mice underwent UUO and were treated with Ang-(1–7) or saline (systemic infusion by osmotic minipumps). (A) A representative western blot showing renal Bax and Bcl-xL expression (upper panel) and the calculation of the Bax/Bcl-xl ratio (lower panel) after 5 days of UUO. Open and black bars represent data of saline- or Ang-(1–7)-infused mice, respectively, expressed as mean±SEM of 6 animals per group. Figures B and C show representative kidney sections (B: cortex with glomerulus; C: medulla) with van Gieson staining and anti-F4/80 immunostaining, respectively (positive immunoreactivity reveals brown signal [arrowheads]). Magnification: 400×. (D) Infiltrating leukocytes were counted based on their morphology and location within renal cortex, medulla, and pelvis in mice infused with saline or Ang-(1–7). (E) Renal NF-κB activation was measured in wild-type mice infused with saline or Ang-(1–7) and in Mas-deficient mice (Mas−/−) and their respective wild-type controls (Mas+/+) at 2 and 5 days of UUO. The figure shows data of renal NF-κB activity expressed as n-fold increase vs. contralateral kidney as mean±SEM of 6 animals per group analyzed in duplicate. (F) Shows a representative EMSA experiment. Competition assay with a 100-fold excess of unlabeled NF-κB shows the specificity of the binding (marked by arrows). The position of free oligonucleotides is indicated. (G) Shows data of gene expression of proinflammatory factors (MCP-1 and IL-6) obtained by real-time PCR experiments and expressed as n-fold increase vs. contralateral kidney as mean±SEM of 8–10 animals per group; C: contralateral, O: obstructed kidneys. * P<0.05 vs. contralateral kidney of the same genotype; # P<0.05 vs. Mas+/+ obstructed kidney; ¥ P<0.05 vs. saline-infused obstructed kidneys.