Table 1.
Terminal kidney function measurements.
Figure 1.
CM treatment increases kidney function.
CM treatment increased glomerular filtration rate (GFR) (A) and effective renal plasma flow (ERPF) (B) in CKD. 2K-NCM (n = 6); 2K-CM (n = 6); CKD-CM (n = 13); CKD-NCM (n = 13). Post-hoc test p-value is shown.
Figure 2.
CM treatment decreased systolic blood pressure (A) and proteinuria (B) in CKD.
2K-NCM (n = 6); ♦ 2K-CM (n = 6); ▪ CKD-CM (n = 13); • CKD-NCM (n = 13). *P<0.05: CKD vs. 2K, † P<0.05: CKD-CM vs. CKD-NCM.
Table 2.
Longitudinal measurements after CM treatment at week 6 after SNX.
Figure 3.
CM treatment increased the number of glomerular endothelial cells (A, C-D), but did not increase tubular endothelial cell number (B, E-F) shown by a JG12 staining.
2K-NCM (n = 6); 2K-CM (n = 6); CKD-NCM (n = 13); CKD-CM (n = 13). *P<0.05: CKD vs. 2K, †P<0.05: CKD-CM vs. CKD-NCM.
Figure 4.
Glomerulosclerosis was reduced by CM treatment.
On PAS-stained sections, glomeurosclerosis was reduced after CM treatment (A). Normal (B), segmental (C) and totally sclerotic are shown (D). Black arrows indicate sclerotic areas. 2K-NCM (n = 6); 2K-CM (n = 6); CKD-NCM (n = 13); CKD-CM (n = 13). *P<0.05: CKD vs. 2K, †P<0.05: CKD-CM vs. CKD-NCM.
Figure 5.
CM treatment increased DNA damage repair.
After logarithmic transformation, more γ-H2AX positive cells, indicating glomerular nuclei undergoing DNA damage repair, were found after CM treatment in CKD (A). Black arrow indicates positive nucleus (B). 2K-NCM (n = 6); 2K-CM (n = 6); CKD-NCM (n = 13); CKD-CM (n = 13). *P<0.05: CKD vs. 2K.
Table 3.
Renal morphology.
Figure 6.
Tubulo-interstitial damage on PAS-stained sections was reduced after CM treatment in CKD rats.
CM treatment decreased tubular fibrosis and atrophy (A). Differences between CKD (B) and healthy (C) tubular kidney tissue are shown. White arrow indicates infitration, black arrows showing proteincasts. 2K-NCM (n = 6); 2K-CM (n = 6); CKD-NCM (n = 13); CKD-CM (n = 13). *P<0.05: CKD vs. 2K, †P<0.05: CKD-CM vs. CKD-NCM.
Figure 7.
Tubular collagen I and III deposition was reduced after CM treatment in CKD rats.
CM treatment reduced sirius red staining (A). Raw (B) and polarized (C) pictures are shown. 2K-NCM (n = 6); 2K-CM (n = 6); CKD-NCM (n = 13); CKD-CM (n = 13). *P<0.05: CKD vs. 2K, †P<0.05: CKD-CM vs. CKD-NCM.
Figure 8.
Inflammatory cytokine expression in healthy and CKD kidneys.
A: Cytokine array in array. Striped bars = 2K-CM (n = 6); dotted bars = 2K-NCM (n = 6); black bars = CKD-NCM (n = 7); white bars = CKD-CM (n = 4). IL-8 = interleukin 8; sICAM = soluble Inter-Cellular Adhesion Molecule; L-selectin = leucocyte cell-adhesion molecule; RANTES = Regulated upon Activation, Normal T-cell Expressed, and Secreted; MIG = Monokine Induced by Gamma-Interferon; MIP1α = Macrophage Inflammatory Protein-1 alpha; MIP3α = Macrophage Inflammatory Protein-3 alpha; Timp-1 = Tissue Inhibitor of Metalloproteinase 1; VEGF = Vascular Endothelial Growth Factor; IL-1RA = Interleukin 1 Receptor Antagonist. B: Fractalkine mRNA in healthy and CKD kidneys. Striped bars = 2K-CM (n = 6); dotted bars = 2K-NCM (n = 6); black bars = CKD-NCM (n = 7); white bars = CKD-CM (n = 4). *P<0.05: CKD vs. 2K.
Figure 9.
CM stimulates in vitro angiogenesis (A) and wound closure (B).
Average tube length was increased after CM treatment compared to NCM (C+D). In a scratch wound assay, CM treatment increased wound (indicated by black arrow in E) closure compared to NCM treatment. *P<0.05 CM vs. NCM.
Figure 10.
Representation of experimental set up.
UNX = uninephrectomy; SNX = subtotal nephrectomy; CM = conditioned medium administration. Stars indicate longitudinal measurements.