Volume 58, Issue 5, Pages 1963-1972 (November 2000) F2-isoprostanes mediate high glucose-induced TGF-β synthesis and glomerular proteinuria in experimental type I diabetes Angel Montero, Karen A. Munger, Rizwan Z. Khan, José M. Valdivielso, Jason D. Morrow, Antonio Guasch, Fuad N. Ziyadeh, Kamal F. Badr Kidney International Volume 58, Issue 5, Pages 1963-1972 (November 2000) DOI: 10.1111/j.1523-1755.2000.00368.x Copyright © 2000 International Society of Nephrology Terms and Conditions
Figure 1 Plasma concentration (A) and urinary excretion (B) of F2-isoprostanes, as determined by gas chromatography/mass spectrometry, in control rats, diabetic rats, and diabetic rats treated with diet supplementation of vitamin E (1000 U/kg diet) four weeks after the onset of diabetes. Symbols in panel B are: (•) diabetes; (♦) diabetes plus vitamin D. *P < 0.05 vs. diabetes; +P < 0.05 vs. control, N = 5 in each group. Time zero values are from control nondiabetic rats. Kidney International 2000 58, 1963-1972DOI: (10.1111/j.1523-1755.2000.00368.x) Copyright © 2000 International Society of Nephrology Terms and Conditions
Figure 2 Daily urinary protein excretion (A) and BUN levels (B) in control rats, diabetic rats (•), and diabetic rats treated with diet supplementation of vitamin E (1000 U/kg diet; ♦) four weeks after the onset of diabetes. *P < 0.05 vs. diabetes, N = 5 in each group and each time point. Time zero values are from control nondiabetic rats. Kidney International 2000 58, 1963-1972DOI: (10.1111/j.1523-1755.2000.00368.x) Copyright © 2000 International Society of Nephrology Terms and Conditions
Figure 3 Expression of TGF-β1 in total protein extracted from glomeruli isolated from control rats, diabetic rats, and diabetic rats treated with diet supplementation of vitamin E (1000 U/kg diet) four weeks after the onset of diabetes, as measured by ELISA. *P < 0.05 vs. control; +P < 0.05 vs. diabetes. N = 5 in each group. Kidney International 2000 58, 1963-1972DOI: (10.1111/j.1523-1755.2000.00368.x) Copyright © 2000 International Society of Nephrology Terms and Conditions
Figure 4 Production of 8-iso-PGF2α in supernatant of murine mesangial cells (A) and rat glomerular endothelial cells (B) after 24 hours of incubation in high ambient glucose, as measured by ELISA. Abbreviations are: NG, normal glucose 100 mg/dL; HG, high glucose 450 mg/dL; NAC, N-acetyl-cysteine 5 mmol/L; Indo, indomethacin 0.1 μmol/L. *P < 0.05 vs. NG; +P < 0.05 vs. HG. Each value is the mean of 12 experiments made in triplicate. Kidney International 2000 58, 1963-1972DOI: (10.1111/j.1523-1755.2000.00368.x) Copyright © 2000 International Society of Nephrology Terms and Conditions
Figure 5 Twenty-four-hour production of 8-iso-PGF2α in supernatant of murine mesangial cells (A) and rat glomerular endothelial cells (B) after 14 days of incubation in high ambient glucose, as measured by ELISA. Abbreviations are: NG, normal glucose 100 mg/dL; HG, high glucose 450 mg/dL; NAC, N-acetyl-cysteine 5 mmol/L for the last 24 hours. *P < 0.05 vs. NG; +P < 0.05 vs. HG. Each value is the mean of 12 experiments made in triplicate. Kidney International 2000 58, 1963-1972DOI: (10.1111/j.1523-1755.2000.00368.x) Copyright © 2000 International Society of Nephrology Terms and Conditions
Figure 6 Production of TGF-β in the supernatant of murine mesangial cells (A) and rat glomerular endothelial cells (B) after 24 hours with 8-iso-PGF2α 0.1 μmol/L, as measured by ELISA. Abbreviations are: Control, cells incubated with vehicle; Iso, cells incubated with 0.01 μmol/L 8-iso-PGF2α. *P < 0.05 vs. control. Each value is the mean of 12 experiments made in triplicate. Kidney International 2000 58, 1963-1972DOI: (10.1111/j.1523-1755.2000.00368.x) Copyright © 2000 International Society of Nephrology Terms and Conditions
Figure 7 Production of TGF-β in supernatant of murine mesangial cells (A) and rat glomerular endothelial cells (B) after 24 hours of incubation in high ambient glucose, as measured by ELISA. Abbreviations are: NG, normal glucose 100 mg/dL; HG, high glucose 450 mg/dL; NAC, N-acetyl-cysteine 5 mmol/L; Indo, indomethacin 0.1 μmol/L. *P < 0.05 vs. NG; +P < 0.05 vs. HG. Each value is the mean of 12 experiments made in triplicate. Kidney International 2000 58, 1963-1972DOI: (10.1111/j.1523-1755.2000.00368.x) Copyright © 2000 International Society of Nephrology Terms and Conditions
Figure 8 Twenty-four hour production of TGF-β in supernatant of murine mesangial cells (A) and rat glomerular endothelial cells (B) after 14 days of incubation in high ambient glucose, as measured by ELISA. Abbreviations are: NG, normal glucose 100 mg/dL; HG, high glucose 450 mg/dL; NAC, N-acetyl-cysteine 5 mmol/L. *P < 0.05 vs. NG; +P < 0.05 vs. HG. Each value is the mean of 12 experiments made in triplicate. Kidney International 2000 58, 1963-1972DOI: (10.1111/j.1523-1755.2000.00368.x) Copyright © 2000 International Society of Nephrology Terms and Conditions
Figure 9 Proposed model summarizing the interactions between isoprostane and TGF-β production by glomerular cells in response to high ambient glucose. While high ambient glucose drives F2-isoprostane formation in both mesangial and endothelial cells, the predominant source of TGF-β is the mesangial cell, on which F2-isoprostanes act in an autocrine and paracrine fashion. Kidney International 2000 58, 1963-1972DOI: (10.1111/j.1523-1755.2000.00368.x) Copyright © 2000 International Society of Nephrology Terms and Conditions