Table 1.
Demographic and clinical characteristics of the patients.
Table 2.
Histopathological findings of the renal biopsies.
Fig 1.
Boxplots of logarithmic transformed medians and 25th to 75th percentiles of the quantification levels of normalized mRNA (2-ΔΔCt) of alpha-actinin-4, podocin, podocalyxin, TGF-β1, CTGF, and VEGF-A kidney tissue expression according to each group of glomerular disease relative to the mRNA (2-ΔΔCt) expression measured in control biopsies.
For each graphic showing the level of podocyte protein or profibrotic factor expression, the p-values as calculated by the Kruskal-Wallis test are presented.
Fig 2.
Immunohistochemistry in kidney biopsies for the histologic assessment of the renal fibrosis by Sirius Red, CTGF, and HHF35 staining.
Sirius Red (unpolarized; all images with magnification x100): (A) Strong red staining of collagen fibers in the interstitium and in the glomeruli. (B) Moderate staining of collagen fibers distributed in the interstitium and a weak staining in the glomeruli. (C) No staining was detected in a control biopsy. CTGF (diaminobenzidine; all images with magnification x100): (D) Moderate to intense staining expressed in podocytes and along the glomerular capillary walls, very strong staining expressed in tubules, and a very weak staining in the interstitium. (E) Moderate staining in the tubules, weak in the glomeruli, and no staining in the interstitium. (F) No staining in the glomeruli and in the interstitium, and a very weak staining in a few tubules detected in a control biopsy. HHF35 (diaminobenzidine, all images with magnification x100): (G) Strong staining of smooth muscle bundles with cellular intimal thickening and intimal fibrosis in the cortical interstitium, and to a lesser degree, in the tubules and in the glomerular capillary walls. (H) Moderate staining in the tubules, interstitium, and in a glomerulus. (I) Absence of or a scarce staining in a few tubules in a control biopsy.
Table 3.
Median and interquartile ranges of the percent positive area of fibrosis in renal biopsies by staining in each glomerular disease in relation to control biopsies and between pairs of glomerular diseases.
Fig 3.
Spearman’s correlations between the percent positive area of fibrosis measured by HHF35, CTGF, and Sirius Red staining with the log transformed mRNA (2-ΔΔCt) of TGF-β1, CTGF and VEGF-A, as expressed in renal biopsies of patients diagnosed with podocytopathies, proliferative nephritis, diabetic kidney disease, and IgA nephropathy.
In each graphic, the correlation coefficient rs and respective p-values are presented. The three rows all have different y-axis scales.
Fig 4.
Spearman’s correlations between the percent positive area of fibrosis measured by CTGF staining and the log transformed mRNA (2-ΔΔCt) of alpha-actinin-4, podocin, and podocalyxin, as expressed in renal biopsies of patients diagnosed with podocytopathies, proliferative nephritis, diabetic kidney disease, and IgA nephropathy.
In each graphic, the correlation coefficient rs and respective p-values are presented.
Fig 5.
Spearman’s correlations between the percentage positive area of fibrosis measured by Sirius Red staining with renal function, as estimated by glomerular filtration rate at the time of renal biopsy (initial) and at last follow up (final).
In each graphic, the correlation coefficient rs and respective p-values are presented.