Volume 58, Issue 3, Pages 1219-1227 (September 2000) Aldosterone modulates plasminogen activator inhibitor-1 and glomerulosclerosis in vivo  Nancy J. Brown, Shinya Nakamura, Lijun Ma, Ikuko Nakamura, Ellen Donnert, Michael Freeman, Douglas E. Vaughan, Agnes B. Fogo  Kidney International  Volume 58, Issue 3, Pages 1219-1227 (September 2000) DOI: 10.1046/j.1523-1755.2000.00277.x Copyright © 2000 International Society of Nephrology Terms and Conditions

Figure 1 Systolic blood pressure (SBP) measured by tail cuff plethysmography 12 weeks following radiation or sham radiation in nonradiated (N), placebo-treated radiated (R + P), spironolactone-treated radiated (R + SP), angiotensin type 1 receptor antagonist (AT1RA)-treated radiated (R + AT1RA), and spironolactone + AT1RA-treated radiated (R + SP + AT1RA) rats. Kidney International 2000 58, 1219-1227DOI: (10.1046/j.1523-1755.2000.00277.x) Copyright © 2000 International Society of Nephrology Terms and Conditions

Figure 2 Twenty-four–hour urine protein excretion 12 weeks following radiation or sham radiation in nonradiated (N), placebo-treated radiated (R + P), spironolactone-treated radiated (R + SP), AT1RA-treated radiated (R + AT1RA), and spironolactone + AT1RA-treated radiated (R + SP + AT1RA) rats. Kidney International 2000 58, 1219-1227DOI: (10.1046/j.1523-1755.2000.00277.x) Copyright © 2000 International Society of Nephrology Terms and Conditions

Figure 3 Sclerosis index assessed in kidneys obtained from placebo-treated (R + P) radiated, spironolactone-treated radiated (R + SP), AT1RA-treated radiated (R + AT1RA), and spironolactone + AT1RA-treated radiated (R + SP + AT1RA) rats sacrificed at 12 weeks. The sclerosis index was calculated as outlined in the text. Kidney International 2000 58, 1219-1227DOI: (10.1046/j.1523-1755.2000.00277.x) Copyright © 2000 International Society of Nephrology Terms and Conditions

Figure 4 Expression of plasminogen activator inhibitor-1 (PAI-1) mRNA, normalized for GAPDH mRNA expression, measured in kidneys obtained from placebo-treated radiated (R + P), spironolactone-treated radiated (R + SP), AT1RA-treated radiated (R + AT1RA), and spironolactone + AT1RA-treated radiated (R + SP + AT1RA) rats sacrificed at 12 weeks. Kidney International 2000 58, 1219-1227DOI: (10.1046/j.1523-1755.2000.00277.x) Copyright © 2000 International Society of Nephrology Terms and Conditions

Figure 5 Photomicrographs of kidney sections from placebo-treated and spironolactone-treated radiated rats. At 12 weeks, the kidney from placebo-treated radiated rats showed moderate tubulointerstitial damage and glomerulosclerosis (A). In spironolactone-treated rats, tubulointerstitial damage and glomerulosclerosis were ameliorated, as illustrated by decreased collagen staining (B; Masson stain A and B, ×100). PAI-1 immunostaining was increased diffusely in the tubulointerstitium and was accentuated in areas of segmental sclerosis in placebo-treated radiated rats (C, ×400). By in situ hybridization (E, ×400) PAI-1 mRNA expression exhibited a similar distribution, with accentuation in sclerotic glomeruli in placebo-treated radiated rats. In areas of injury, PAI-1 mRNA signal was present in mesangial areas and glomerular visceral epithelial cells (arrows). There was no signal after in situ hybridization with sense probes (data not shown). In the spironolactone-treated rats, PAI-1 immunostaining (D, ×200) and mRNA (F, ×200) were decreased in both the tubular interstitium and in glomeruli (arrow). By in situ hybridization, transforming growth factor-β (TGF-β) mRNA was mildly and diffusely increased with occasional accentuation in the glomeruli of placebo-treated radiated rats (G, ×400). TGF-β mRNA was mildly attenuated by treatment with spironolactone (H, ×400). Kidney International 2000 58, 1219-1227DOI: (10.1046/j.1523-1755.2000.00277.x) Copyright © 2000 International Society of Nephrology Terms and Conditions

Figure 6 Relationship between the sclerosis index and plasminogen activator inhibitor 1 (PAI-1) immunostaining for glomeruli from all radiated rats. Symbols are: (•) N = 1; (▴) N = 10; (▪) N = 100. Kidney International 2000 58, 1219-1227DOI: (10.1046/j.1523-1755.2000.00277.x) Copyright © 2000 International Society of Nephrology Terms and Conditions