Figure 1.
Adipocyte and osteogenic differentiation and MSC tracking assay.
Standard culture (A), differentiated adipocytes containing lipid droplets visualized by Oil Red coloration (B), and osteocytes exhibiting calcium deposition (C), demonstrated by Alizarin coloration. Magnification: 100x. (D) MSC immunophenotyping. MSCs were isolated and labeled for CD73, CD90, CD44 and CD34. Labeled MSC with fluorescent marker detected by cytometry of 2K-1C animals (n = 2) and 2K-1C + MSC (n = 2) in the cortex and medulla of the clipped kidney (E), unclipped kidney (F), lung (G) and medulla oblongata (H). Percentage of labeled cells found in the cortex and medulla of clipped and unclipped kidney, lung and medulla oblongata, analyzed by FACS (I).
Figure 2.
Systolic blood pressure (SBP), renal sympathetic nerve activity (RSNA) and renal functional parameters.
(A) SBP expressed as the mean ± SEM. *P<0.05 vs. SHAM. +P<0.05 vs. 2K-1C (2-way ANOVA followed by the Bonferroni’s post-test). RSNA (B), plasma creatinine (C), proteinuria (D) and body weight (E) of SHAM animals (n = 5), 2K-1C animals (n = 7) and 2K-1C + MSC animals (n = 7). *P<0.05 vs. SHAM. +P<0.05 vs. 2K-1C. (One-way ANOVA followed by the Newman Keuls’s post-test).
Table 1.
Plasma and urinary parameters.
Figure 3.
Morphology and fibrosis of the kidney.
Hematoxylin-Eosin (H&E) (A-F) and Picrosirius red (G-L) staining (20x) of a renal cortex and medulla of the clipped kidneys of SHAM, 2K-1C and 2K-1C + MSC animals. Fibrotic area was measured using the Image Tool program (M and N). *P<0.05 vs. 2K-1C. (One-way ANOVA followed by the Newman Keuls’s post-test.
Figure 4.
Representative image of JG-12 staining of a renal cortex (20x) and medulla (40X) of the clipped kidney of a SHAM (A and D), 2K-1C (B and E) and 2K-1C + MSC animal (C and F). Stained areas were measured using the Image Tool program (G and H). The arrows show the positive staining in the arterioles. *P<0.05 vs. SHAM. +P<0.05 vs. 2K-1C (One-way ANOVA followed by the Tukey’s post-test).
Figure 5.
Expression of components of Renin-Angiotensin (RAS) in the medulla of clipped kidneys.
Angiotensinogen (A), Renin (B), ACE (C), AT1 (D) and AT2 receptors (E) in the medulla of kidney clipped of SHAM (n = 5), 2K-1C (n = 7) and 2K-1C + MSC animals (n = 7). *P<0.05 vs. SHAM. +P<0.05 vs. 2K-1C (One-way ANOVA followed by the Tukey’s post-test).
Figure 6.
(A) IL-1β mRNA expression. (B) TNF-α mRNA expression. (C) IL-10 mRNA expression. Data expressed as arbitrary units *P<0.05 vs. SHAM. +P<0.05 vs. 2K-1C. (One-way ANOVA followed by the Tukey’s post-test).