Figure 6

Burns septic acute renal failure (ARF) group plasma altered polarity, permeability to albumin and nephrin expression on podocytes. (a) Significant variation of trans-epithelial resistance (TER) after exposure to burns septic ARF group plasma (*p < 0.05 burns septic ARF group vs control healthy plasma). (b-d) Representative immunofluorescence images of the altered distribution of the intermediate filament protein nestin in presence of burns septic plasma (d), not detectable after incubation with vehicle alone (b) or control healthy plasma (c) (magnification × 400, nuclei counterstained with 1 μg/ml propidium iodide). After burns septic plasma challenge, nestin lost its typical diffuse distribution and was localized in the sub-membrane spaces forming several "rings" (d). (e) Significant increased diffusion of albumin across podocyte monolayers exposed to burns septic ARF group plasma (*p < 0.05 burns septic plasma vs control healthy plasma). (f-h) Representative immunofluorescence images of the slit diaphragm protein nephrin after exposure to vehicle alone (f), control healthy plasma (g) and burns septic ARF group plasma (h). Burns septic ARF group plasma induced a diffuse loss of nephrin (magnification × 400, nuclei counterstained with 1 μg/ml propidium iodide). (i) Representative Western blot analysis of podocyte nephrin expression (vehicle alone in lane 1, control healthy plasma in lanes 2–5, burns septic ARF group plasma in lanes 6–9) and related densitometric analysis. Beta-actin was used as reference for protein loading. Values in (a) and (e) are expressed as averages ± standard error (SE). Each plasma was tested in triplicate. Analysis of variance (ANOVA) with Newman-Keuls multi-comparison test was performed.