Apoptosis in the kidneys of patients with type II diabetic nephropathy D. Verzola, M.T. Gandolfo, F. Ferrario, M.P. Rastaldi, B. Villaggio, F. Gianiorio, M. Giannoni, L. Rimoldi, F. Lauria, M. Miji, G. Deferrari, G. Garibotto  Kidney International  Volume 72, Issue 10, Pages 1262-1272 (November 2007) DOI: 10.1038/sj.ki.5002531 Copyright © 2007 International Society of Nephrology Terms and Conditions

Figure 1 Glomerular and tubular apoptosis by ssDNA antibodies analysis. Serial sections of kidneys with type II diabetic nephropathy were stained for apoptosis using the ssDNA Ab. Black arrows point to cells that display the nuclear fragmentation and condensation characteristic of apoptosis. The anti-ssDNA signal was not only confined to glomerular cells (a and b, mesangial cells; c, podocytes; d, endothelial cells), but it was also detected (e, f) in tubular and (g, h) vascular endothelial cells (original magnification × 1000). Kidney International 2007 72, 1262-1272DOI: (10.1038/sj.ki.5002531) Copyright © 2007 International Society of Nephrology Terms and Conditions

Figure 2 Apoptosis in the kidneys of patients with type II diabetic nephropathy. (a) Patients with type II diabetic nephropathy (DN) presented an increase in apoptotic index both in the glomeruli and tubuli when compared to controls and patients affected by nondiabetic nephropathies (GN). (b) Apoptosis was similarly enhanced both in patients with early and in those with advanced clinical impairment. aP<0.05; bP<0.01 vs controls and patients with GN and proteinuria. Mean±s.e.m. Kidney International 2007 72, 1262-1272DOI: (10.1038/sj.ki.5002531) Copyright © 2007 International Society of Nephrology Terms and Conditions

Figure 3 Cell fate in type II diabetic nephropathy. Apoptosis and proliferation were quantified in the glomeruli and tubules in kidney biopsies from patients with type II DN. In these patients, the proliferation/apoptosis ratio was markedly decreased in favor of apoptosis both in glomeruli and tubuli. aP<0.05; bP<0.01 vs controls. Mean±s.e.m. Kidney International 2007 72, 1262-1272DOI: (10.1038/sj.ki.5002531) Copyright © 2007 International Society of Nephrology Terms and Conditions

Figure 4 Apoptosis in the kidneys of patients with type II diabetic nephropathy. Apoptosis was directly associated with HbA1c levels in the (a) glomeruli but not in (b) tubule cells in patients with type II diabetic nephropathy. Kidney International 2007 72, 1262-1272DOI: (10.1038/sj.ki.5002531) Copyright © 2007 International Society of Nephrology Terms and Conditions

Figure 5 Apoptosis in the kidneys of patients with type II diabetic nephropathy. Apoptosis was directly associated with systolic blood pressure in the (a) glomeruli but not in (b) tubule cells in patients with type II diabetic nephropathy. Kidney International 2007 72, 1262-1272DOI: (10.1038/sj.ki.5002531) Copyright © 2007 International Society of Nephrology Terms and Conditions

Figure 6 Apoptosis in the kidneys of patients with type II diabetic nephropathy. Apoptosis was associated with plasma LDL-cholesterol in (b) tubule but not in (a) glomerular cells in patients with type II diabetic nephropathy. Kidney International 2007 72, 1262-1272DOI: (10.1038/sj.ki.5002531) Copyright © 2007 International Society of Nephrology Terms and Conditions

Figure 7 Expression of Fas (immunohistochemistry) in (a–c) controls and in (d–f) kidney biopsies from patients with type II diabetic nephropathy (c–f: original magnification × 400; a, b: original magnification × 1000). Results of image analysis are shown in g and h (aP<0.05 vs controls). Fas expression was clearly upregulated both in glomeruli and tubuli. Mean±s.e.m. Kidney International 2007 72, 1262-1272DOI: (10.1038/sj.ki.5002531) Copyright © 2007 International Society of Nephrology Terms and Conditions

Figure 8 Expression of FasL (immunohistochemistry and immunofluorescence) in (a, d) controls and in (b, c, e, f, h, i) patients with type II diabetic nephropathy. FasL was upregulated both in the glomeruli and tubuli in patients with diabetic nephropathy. (j, k) Image analysis (aP<0.05, bP<0.01 vs controls) (b, g, h: original magnification × 400; a, d, f, i: original magnification × 1000). Mean±s.e.m. Kidney International 2007 72, 1262-1272DOI: (10.1038/sj.ki.5002531) Copyright © 2007 International Society of Nephrology Terms and Conditions

Figure 9 Relationships between Fas/FasL expression in the kidney and estimated GFR. FasL expression was inversely related to estimated GFR in (b) the glomeruli, but not in (d) tubuli, in patients with type II diabetic nephropathy. (a) Glomerular and (c) tubular Fas expression was not related to estimated GFR. Kidney International 2007 72, 1262-1272DOI: (10.1038/sj.ki.5002531) Copyright © 2007 International Society of Nephrology Terms and Conditions

Figure 10 Expression of phosphorylated p38 MAPK (immunohistochemistry) in (a) controls (original magnification × 400) and in (b–d) patients with type II diabetic nephropathy (original magnification × 1000). Phosphorylated p38 was upregulated in glomerular, tubular (aP<0.05 vs controls), and vascular endothelial cells (a: original magnification × 400; b–d: original magnification × 1000). Data±s.e.m. Kidney International 2007 72, 1262-1272DOI: (10.1038/sj.ki.5002531) Copyright © 2007 International Society of Nephrology Terms and Conditions

Figure 11 Relationships between (a) glomerular and (b) tubule cell apoptosis and loss of renal function at 2 years in patients with type II diabetic nephropathy and proteinuria. Kidney International 2007 72, 1262-1272DOI: (10.1038/sj.ki.5002531) Copyright © 2007 International Society of Nephrology Terms and Conditions