Organoprotective Effects of Spironolactone on Top of Ramipril Therapy in a Mouse Model for Alport Syndrome

Figure 1. Serum aldosterone levels indicating an aldosterone-escape in Alport mice receiving Angiotensin-converting enzyme inhibitor (ACEi)-monotherapy.

Figure 2. Effect on protein excretion and kidney function on days 50, 75 and 100 of therapy with ACEi and ACEi+Spiro. (A) Total renal protein excretion (relative to untreated mice at 65 days) in the ACEi group compared to the ACEi+Spiro group (n = 3 for each group); (B) Relative change in excretion of high molecular weight proteins (glomerular damage), albumin and low molecular weight proteins (tubular damage) in the ACEi group compared to the ACEi+Spiro group (n = 3 for each group); (C) Blood urea nitrogen (BUN) levels in the untreated mice compared to the ACEi group and the ACEi+Spiro group (n = 3 for WT and ACEi 100d, n = 4 for Plac, ACEi 75d and ACEi+Spiro 75d, n = 2 for ACEi+Spiro 100d); *** p < 0.001; ** p < 0.01; * p < 0.05.

Figure 3. Therapeutic effect on matrix accumulation during pathogenesis of Alport Syndrome (AS). Immune-fluorescence staining of laminin 111, magnification 100-fold. (A) wildtype control; (B) Alport mice, ACEi 75 days; (C) Alport mice, ACEi 100 days; (D) Alport mice, Plac (untreated); (E) Alport mice, ACEi+Spiro 75 days; (F) Alport mice, ACEi-Spiro 100 days; (G) score for tubulointerstitial matrix accumulation (n = 3 for each group); (H) score for glomerular matrix accumulation (n = 3 for each group); *** p < 0.001; ** p < 0.01; * p < 0.05.

Figure 4. Therapeutic effect on scar tissue formation during pathogenesis of AS. Immune-fluorescence staining of fibronectin, magnification 200-fold. (A) wildtype control; (B) Alport mice, ACEi 75 days; (C) Alport mice, ACEi 100 days; (D) Alport mice, Plac (untreated); (E) Alport mice, ACEi+Spiro 75 days; (F) Alport mice, ACEi-Spiro 100 days; (G) score of tubulointerstitial fibrosis (n = 3 for each group); (H) score of glomerulosclerosis and fibrosis (n = 3 for each group); *** p < 0.001; ** p < 0.01; * p < 0.05.

Figure 5. Therapeutic effect on timespan until end-stage renal failure. (A) Kaplan–Meier curves of lifespan in days until end-stage renal failure in untreated (Plac), ACEi treated and Alport mice with dual RAAS-blockade; (B) Box-blot of lifespan in untreated (Plac), ACEi treated and ACEi+Spiro treated Alport mice. n = 8 for Plac, n = 10 for ACEi and n = 10 for ACEi+Spiro; *** p < 0.001; * p < 0.05.

Figure 6. Anti-androgen side-effects of spironolactone on body weight. (A) Mean body weight of male and female Alport mice in the ACEi group vs. dual treated mice (ACEi+Spiro); (B) Individual measures of body weight of fully grown adult male mice in the ACEi group vs. dual treated males (ACEi+Spiro) as shown in the box in graph A (n = 10–13 datapoints for each group). m = male; f = female; ** p < 0.01.