Volume 62, Issue 4, Pages 1349-1357 (October 2002) Vasopeptidase inhibition and Ang-(1-7) in the spontaneously hypertensive rat  Carlos M. Ferrario, David B. Averill, K. Bridget Brosnihan, Mark C. Chappell, Samy S. Iskandar, Richard H. Dean, Debra I. Diz  Kidney International  Volume 62, Issue 4, Pages 1349-1357 (October 2002) DOI: 10.1111/j.1523-1755.2002.kid559.x Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 1 Average changes in tail-cuff systolic blood pressure.(A) Vehicle (▪) and (B) omapatrilat-treated SHR (□) during a 17-day dosing period. Values are means ± SE. *P < 0.01 compared to vehicle. Kidney International 2002 62, 1349-1357DOI: (10.1111/j.1523-1755.2002.kid559.x) Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 2 Omapatrilat causes a significant increase in plasma concentrations of angiotensins [Ang I, Ang II, and Ang-(1-7)] at day 17 of treatment period. Values are means ± SE of SHR given either vehicle (□) or omapatrilat (▪). *P < 0.02; **P < 0.001. Kidney International 2002 62, 1349-1357DOI: (10.1111/j.1523-1755.2002.kid559.x) Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 3 Effect of chronic exposure to omapatrilat (□) compared to vehicle-treated controls (▪).(A) cumulative water intake. (B) Urine volume. (C) urine osmolality. (D) urinary sodium excretion. (E) Urinary potassium excretion. (F) Food consumption. Values are means ± SE; *P < 0.05 compared to vehicle. Kidney International 2002 62, 1349-1357DOI: (10.1111/j.1523-1755.2002.kid559.x) Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 4 Excretion of angiotensins [Ang I(A), Ang II(B), and Ang-(1-7)(C)] is markedly augmented throughout the 17-day duration of exposure. Symbols are: omapatrilat (□); SHR given vehicle (▪). Values are means ± SE; *P < 0.01 compared to vehicle. Kidney International 2002 62, 1349-1357DOI: (10.1111/j.1523-1755.2002.kid559.x) Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 5 Comparison of absorbance using high pressure liquid chromatography.(A) High pressure liquid chromatography (HPLC) of angiotensin-(1-7) [Ang-(1-7)] metabolism in dialyzed rat urine for 15 minutes at 37°C reveals conversion to Ang-(1-4) and Ang-(1-5) [control, solid lines]. Addition of one μmol/L omapatrilat [OMAP, dashed lines] attenuated generation of these angiotensin metabolites and increased the recovery of Ang-(1-7). (B) Time course for the hydrolysis of Ang-(1-7) at 37°C in rat urine alone (○) or in the presence of omapatrilat (•). Kidney International 2002 62, 1349-1357DOI: (10.1111/j.1523-1755.2002.kid559.x) Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 6 Photomicrographs of sections of kidneys. SHR given either vehicle (A) and (C) or omapatrilat (B) and (D) for 17 days. (A) and (C) from a vehicle-treated SHR shows that Ang-(1-7) immunostaining is present in renal proximal convoluted tubules throughout the cortex while the medulla showed scant Ang-(1-7)-positive staining cells. No immunopositive Ang-(1-7) staining is found in glomeruli. (B) shows an apparent increase in the density of Ang-(1-7) immunostaining in renal proximal tubules, including increased immunostaining now also found in the thick portion of the ascending limb of Henle's loops (D). Magnification for (A) and (B) ×2; magnification for (C) and (D) ×100. Kidney International 2002 62, 1349-1357DOI: (10.1111/j.1523-1755.2002.kid559.x) Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 7 Preabsorption of the antibody with 10 μmol/L Ang-(1-7). Complete elimination of staining is shown in an adjacent section from a vehicle treated SHR. (A) Magnification ×25; (B) Magnifcation ×20. Kidney International 2002 62, 1349-1357DOI: (10.1111/j.1523-1755.2002.kid559.x) Copyright © 2002 International Society of Nephrology Terms and Conditions