Combining Neprilysin Inhibitor With AT2R Agonist Is Superior to Combination With AT1R Blocker in Providing Reno-Protection in Obese Rats

Figures

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  FIGURE 1. (A) kidney weight, (B) urinary protein normalized to 24 h urine volume, (C) urinary albumin normalized to 24 h urine volume in obese Zucker rats on NSD or HSD and treated with vehicle, SAC/C21, or SAC/VAL. Data shown as mean ± SEM, analyzed by one-way ANOVA, followed by Fisher’s LSD test. Results are considered significant at *p < 0.05, with a 95% confidence interval; n = 8. NSD—normal salt diet fed + vehicle treated obese control, HSD—high salt diet fed + vehicle treated obese control, SAC/C21—high salt diet fed obese rat treated with sacubitril + C21, SAC/VAL—high salt diet fed obese rat treated with sacubitril + valsartan.

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  FIGURE 2. Analysis of renal injury via markers and indicators of glomerular and tubular damage (A) urinary osteopontin normalized to 24 h urine volume, (B) urinary cystatin C normalized to 24 h urine volume, (C) urinary creatinine, (D) plasma cystatin C, (E) plasma creatinine, (F) eGFR, (G) nephrin expression in kidney cortex, (H) podocin expression in kidney cortex, and (I) megalin expression in kidney cortex from obese Zucker rats on NSD or HSD and treated with vehicle, SAC/C21, or SAC/VAL. Data shown as mean ± SEM, analyzed by one-way ANOVA, followed by Fisher’s LSD test. Results are considered significant at *p < 0.05, **p < 0.01, ****p < 0.0001 with a 95% confidence interval; n = 6–8 for urinary osteopontin, plasma creatinine, and eGFR and n = 8 for urinary and plasma cystatin C, urinary creatinine, nephrin, podocin, and megalin expression. NSD—normal salt diet fed + vehicle treated obese control, HSD—high salt diet fed + vehicle treated obese control, SAC/C21—high salt diet fed obese rat treated with sacubitril + C21, SAC/VAL—high salt diet fed obese rat treated with sacubitril + valsartan.

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  FIGURE 3. (A) plasma ANP, (B) Neprilysin activity in kidney cortex, (C) Neprilysin expression in kidney cortex, (D) NPR-C expression in kidney cortex, (E) NPR-C expression in white adipose tissue, (F) plasma bradykinin, (G) ACE2 expression in kidney cortex, and (H) ACE2 activity in kidney cortex from obese Zucker rats on NSD or HSD and treated with vehicle, SAC/C21, or SAC/VAL. Data shown as mean ± SEM, analyzed by one-way ANOVA, followed by Fisher’s LSD test. Results are considered significant at *p < 0.05, with a 95% confidence interval; n = 6–8 for neprilysin activity and renal NPR-C and ACE2 expression and n = 8 for plasma ANP and bradykinin, expression of renal neprilysin and adipose NPR-C, and renal ACE2 activity. NSD—normal salt diet fed + vehicle treated obese control, HSD—high salt diet fed + vehicle treated obese control, SAC/C21—high salt diet fed obese rat treated with sacubitril + C21, SAC/VAL—high salt diet fed obese rat treated with sacubitril + valsartan.

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  FIGURE 4. Angiotensin peptides, receptors and downstream signaling markers in obese Zucker rats on NSD or HSD and treated with vehicle, SAC/C21, or SAC/VAL. (A) cortical Ang II, (B) renin activity in kidney cortex, (C) plasma renin, (D) AT1R expression in kidney cortex, (E) AT2R expression in kidney cortex from obese Zucker rats on NSD or HSD and treated with vehicle, SAC/C21, or SAC/VAL. Data shown as mean ± SEM, analyzed by one-way ANOVA, followed by Fisher’s LSD test. Results are considered significant at *p < 0.05, with a 95% confidence interval; n = 6–8 for Ang II, plasma renin and AT2R expression and n = 8 renin activity and AT1R expression. NSD—normal salt diet fed + vehicle treated obese control, HSD—high salt diet fed + vehicle treated obese control, SAC/C21—high salt diet fed obese rat treated with sacubitril + C21, SAC/VAL—high salt diet fed obese rat treated with sacubitril + valsartan.