Chronic Metabolic Acidosis Elicits Hypertension via Upregulation of Intrarenal Angiotensin II and Induction of Oxidative Stress

Figures

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  Figure 1. Blood pressure measurement (tail-cuff). Each bar represents values expressed as mean ± SEM. The change in mean arterial pressure (MAP) was considered significant (*) when p < 0.05 between two groups. The data were analyzed using two-way ANOVA followed by Bonferroni posttests.

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  Figure 2. Blood (A) and urine (B) pH measurements. Each bar represents values expressed as mean ± SEM. The change in pH was considered significant (*) when p < 0.05 between two groups. The data were analyzed using two-way ANOVA followed by Bonferroni posttests. Measurement of blood pH (A) and urine pH (B) in control and chronic metabolic acidosis (CMA) animals.

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  Figure 3. Blood pressure measurements for treatment groups (tail-cuff). Each plot represents values expressed as mean ± SEM. The change in MAP was considered significant (*) when p < 0.05 (control vs. CMA) groups and (#) p < 0.05 (CMA vs. CMA + captopril or CMA + tempol) groups. The data were analyzed using two-way ANOVA followed by Bonferroni multiple comparison test among all five groups.

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  Figure 4. Inline-pressure transducer readings for MAP and heart rate. Each bar represents values expressed as mean ± SEM. (A) The change in MAP was considered significant when * p < 0.05 (control vs CMA) groups and # p < 0.05 (CMA vs. CMA + captopril or CMA + tempol) groups. (B) No significant changes in heart rates among the groups. The data were analyzed using one-way ANOVA followed by Tukey’s multiple comparison test among all five groups. t-test was performed to compare two individual groups.

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  Figure 5. Urinary sodium analysis. Each bar represents values expressed as mean ± SEM. The change in urine sodium levels were considered significant (*) when p < 0.05 between groups. The data were analyzed using one-way ANOVA followed by Tukey’s multiple comparison test among all five groups.

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  Figure 6. Serum sodium and potassium levels. Each bar represents values expressed as mean ± SEM. (A) The changes in serum sodium levels were considered significant when * p < 0.05 (control vs. CMA) groups and # p < 0.05 (CMA vs. CMA + captopril or CMA + tempol) groups. (B) The changes in serum potassium levels were significant when * p < 0.05 (Control vs. CMA) groups. The data were analyzed using one-way ANOVA followed by Tukey’s multiple comparison test among all five groups.

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  Figure 7. Interstitial fluid analysis for Angiotensin II concentration. Each bar represents values expressed as mean ± SEM. The changes in interstitial Ang II concentrations were considered significant when * p < 0.05 (control vs. CMA) groups and # p < 0.05 (CMA vs. CMA + captopril or CMA + tempol) groups. The data were analyzed using one-way ANOVA followed by Tukey’s multiple comparison test among all five groups.

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  Figure 8. Plasma aldosterone (A) and plasma renin (B) analysis. Each bar represents values expressed as mean ± SEM. The changes in plasma aldosterone and renin were considered significant when * p < 0.05 (control vs. CMA) groups. The data were analyzed using one-way ANOVA followed by Tukey’s multiple comparison test among all five groups.

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  Figure 9. Periodic acid-Schiff (PAS) staining of kidney cortex sections.

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  Figure 10. The protein expression of Na+/K+ ATPase in the renal proximal tubule. Each bar represents values expressed as mean ± SEM. The changes in protein concentrations were considered significant (*) when p < 0.05 (control vs. CMA) groups and (#) p < 0.05 (CMA vs. CMA + captopril or CMA + tempol) groups. The data were analyzed using one-way ANOVA followed by Tukey’s multiple comparison tests among all five groups.

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  Figure 11. Electron paramagnetic resonance (EPR) analysis for the detection of superoxide and peroxynitrite free radicals. (A,B) show the EPR signal intensities and relative EPR signal area quantification (arbitrary units), respectively, for superoxide radicals.

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  Figure 12. EPR analysis for peroxynitrite. (A,B) show the EPR intensities and the quantified EPR signal area of the first derivative signals, respectively, for peroxynitrite.