Real-time RT-PCR analysis of retinal expression of inflammatory cytokines: VEGF, MCP1, ICAM1 and TNFα.

<p>Values on y-axis represent fold change in mRNA levels compared to WT (normalized as 1). *P<0.01 (vs. DM Ren2+vehicle group). WT, wildtype, DM/NDM, diabetic/nondiabetic. N = 5.</p

Primers used for Real-Time RT-PCR analysis.

<p>Primers used for Real-Time RT-PCR analysis.</p

In situ cell death detection in the retina.

<p>A, Apoptotic cells were detected in-diabetic (NDM) Ren2 rats (a & b), diabetic (DM) Ren2 rats treated with vehicle (c & d), and aliskiren (e & f). ONL: outer nuclear layer; INL: inner nuclear layer; RGC: retinal ganglion cell layer. B, Quantitative measurement of apoptotic cells detected by TUNEL assay. #p<0.01 (vs. wildtype (WT) Sprague–Dawley rats); $p<0.01 (vs. NDM Ren2 and WT); *p<0.01 (vs. NDM Ren2 and DM Ren2+ vehicle).</p

Immunofluorescence detection of GFAP expression in the retinas of wildtype (WT) Sprague–Dawley rats, non-diabetic (NDM) Ren2, diabetic (DM) Ren2 rats treated with vehicle, aliskiren or aliskiren+HRP.

<p>ONL: outer nuclear layer; OPL: outer plexiform layer; INL: inner nuclear layer; IPL: inner plexiform layer; RGC: retinal ganglion cell layer.</p

Representative retinal micrographs of wildtype (WT) Sprague–Dawley rats, non-diabetic (NDM) Ren2 rats, and diabetic (DM) Ren2 rats treated with vehicle, aliskiren or aliskiren+HRP (A).

<p>INL: inner nuclear layer; IPL: inner plexiform layer; RGC: retinal ganglion cell layer. Quantitative measurement of cell density in RGC layer in the central (B) and peripheral (C) retinas of wildtype (WT) Sprague–Dawley rats, non-diabetic (NDM) Ren2 rats, and diabetic (DM) Ren2 rats treated with vehicle, aliskiren or aliskiren+HRP. N = 4. *p<0.01 (vs. WT).</p

Real-time RT-PCR analysis of the expression of inflammatory cytokines IL-1α and TNF-α in cultured Müller cells.

<p>Values on y-axis represent fold change in mRNA levels compared to control (non-treated, normalized as 1). *P<0.01 (vs. control); # P<0.01 (vs. prorenin treated group). N = 4.</p

Early stages of atherogenic DM leads to renal damage.

<p>A: Representative illustration of PAS stained glomeruli from a DM+ATH pig, showing mesangial proliferation and matrix expansion with capillary loops lying around the mesangium as a corona, reminiscent of a beginning Kimmelstiel-Wilson nodule (left panel; thin black arrow). Dilated capillary loops with red cell fragments show intense PAI-1 staining on consecutive slides (right panel; thick black arrow). B: Mesangial expansion index in Controls (n = 7), ATH (n = 5) and DM+ATH (n = 5) pigs. C. Electron microscopy images illustrating a normal GBM architecture (left panel; thick arrow) of the Controls pig. In ATH, there is slight effacement of the podocyte pedicles (middle panel; thick arrow). In DM+ATH, marked lipid deposits were found (right panel). Data are shown as mean ± SEM. *P<0.05 compared to Controls or ATH pigs. Original magnification of A: x400 and C: x8000.</p

No difference in renal vWf and VEGF-A expression.

<p>A. Representative illustrations of kidney sections stained with endothelial marker vWF (arrow: glomerulus; arrowhead: peritubular area) in Controls, ATH, and DM+ATH pigs. B. Representative images of kidney sections stained with VEGF in Controls, ATH, and DM+ATH pigs showing expression in podocytes (arrow head), parietal epithelial cells (thin arrow) and tubuli (asterix). Original magnification of A: x 200 and B: x400.</p

Model characteristics at 15 months of follow up.

<p>Model characteristics at 15 months of follow up.</p

Correlation between capillary tortuosity and Angpt2/Angpt1balance and creatinine levels.

<p>Scatter plot showing the correlation of renal protein expression of Angpt1(A), Angpt2 (B), Angpt2/Angpt1ratio (C).</p