Volume 63, Issue 1, Pages (January 2003)

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Volume 63, Issue 1, Pages 43-52 (January 2003) Therapeutic benefit of spironolactone in experimental chronic cyclosporine A nephrotoxicity  Iris Feria, Israel Pichardo, Patricia Juárez, Victoria Ramírez, Marco A. González, Norma Uribe, Romeo García-Torres, Fernando López- Casillas, Gerardo Gamba, Norma A. Bobadilla  Kidney International  Volume 63, Issue 1, Pages 43-52 (January 2003) DOI: 10.1046/j.1523-1755.2003.00707.x Copyright © 2003 International Society of Nephrology Terms and Conditions

Figure 1 (A) Systolic blood pressure, (B) urinary protein excretion, (C) serum potassium, and (D) serum aldosterone levels in the vehicle group (□), CsA treated animals (), SPIRO group () and CsA+SPIRO treated rats (▪). *P < 0.05 vs. V, SPIRO. Although not shown, serum aldosterone levels in rat fed with normal salt diet are around 100 pg/mL. Kidney International 2003 63, 43-52DOI: (10.1046/j.1523-1755.2003.00707.x) Copyright © 2003 International Society of Nephrology Terms and Conditions

Figure 2 Glomerular filtration rate estimated by creatinine clearance in vehicle group (□), animals treated with CsA during 21 days with 30 mg/kg (), SPIRO group () and CsA+SPIRO treated rats (▪) maintaining on low sodium diet, *P < 0.05 vs. V, SPIRO, and CsA+SPIRO groups. Kidney International 2003 63, 43-52DOI: (10.1046/j.1523-1755.2003.00707.x) Copyright © 2003 International Society of Nephrology Terms and Conditions

Figure 3 (A) Afferent arteriolopathy in CsA treated rat during 21 days (PAS stain ×350). (B) Percentage of afferent arterioles injured, which was estimated in at least 100 preglomerular afferent arterioles from vehicle group (□), CsA treated animals (), SPIRO group () and CsA+SPIRO treated rats (▪). *P < 0.05 vs. V, SPIRO and CsA+SPIRO groups, and †P < 0.05 vs. V, SPIRO and CsA groups. Kidney International 2003 63, 43-52DOI: (10.1046/j.1523-1755.2003.00707.x) Copyright © 2003 International Society of Nephrology Terms and Conditions

Figure 4 (A) Tubulointerstitial fibrosis observed in a salt depleted rat treated with CsA (Van Gieson stain ×200). (B) Mean percentage of injured area obtained in at least ten fields. *P < 0.05 vs. V, SPIRO and CsA+SPIRO groups and †P < 0.05 vs. V, SPIRO and CsA groups. Kidney International 2003 63, 43-52DOI: (10.1046/j.1523-1755.2003.00707.x) Copyright © 2003 International Society of Nephrology Terms and Conditions

Figure 5 Northern blot analysis of mRNA from renal cortex of the four study groups using specific probes for TGF-β1, collagen I, collagen IV, fibronectin and GAPDH. Each lane was loaded with 20 μg of total RNA from a different rat, corresponding to each group as stated. Kidney International 2003 63, 43-52DOI: (10.1046/j.1523-1755.2003.00707.x) Copyright © 2003 International Society of Nephrology Terms and Conditions

Figure 6 TGF-β1 mRNA levels in renal cortex are represented as the ratio between TGF-β1 and GAPDH bands densitometry in vehicle group (□), CsA-treated animals (), SPIRO group () and CsA+SPIRO treated rats (▪).*P < 0.05 vs. V, SPIRO and CsA+SPIRO groups and †P < 0.05 vs. CsA group. Kidney International 2003 63, 43-52DOI: (10.1046/j.1523-1755.2003.00707.x) Copyright © 2003 International Society of Nephrology Terms and Conditions

Figure 7 Effects of CsA and SPIRO on the expression of extracellular matrix proteins. (A) Collagen I, (B) fibronectin and (C) collagen IV in vehicle group (□), CsA-treated animals (), SPIRO group () and CsA+SPIRO treated rats (▪). *P < 0.05 vs. V, SPIRO and CsA+SPIRO groups and †P < 0.05 vs. CsA-treated rats. Kidney International 2003 63, 43-52DOI: (10.1046/j.1523-1755.2003.00707.x) Copyright © 2003 International Society of Nephrology Terms and Conditions

Figure 8 Epidermal growth factor mRNA levels in renal cortex assessed by RT-PCR. (A) An acrylamide gel showing EGF and GAPDH amplification bands for all of the study groups. Each lane was loaded with 10 μL of the RT-PCR reaction from different animals. (B) Graphic representation of the mean ratio between EGF and GAPDH for the Vehicle group (□), CsA-treated animals (), SPIRO group () and CsA+SPIRO treated rats (▪). *P < 0.05 vs. V and †P < 0.05 vs. CsA group. Kidney International 2003 63, 43-52DOI: (10.1046/j.1523-1755.2003.00707.x) Copyright © 2003 International Society of Nephrology Terms and Conditions