Impact of Total or Afferent Renal-Nerve Denervation on Cardiovascular and Renal Function in Sprague Dawley Rats Fed High-Fructose and High-Sodium Diet

Figures

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  Figure 1. Schematic representation of study protocol and timeline.

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  Figure 2. Renal norepinephrine (NE) content in male (A) and female (B) rats. Values for left and right kidneys of G-Sham and F-Sham were combined. Values are mean ± SE. * p < 0.01, ** p < 0.001 and indicated; † p < 0.01 vs. all other groups, § p < 0.001 vs. all groups except F-Sham by ANOVA. (C) Representative renal pelvicalyceal immuno-histochemical staining for CGRP in male rats. (D) Representative renal pelvicalyceal immuno-histochemical staining in female rats. G-Sham, 20% glucose + 0.4% Na; F-Sham, 20% fructose + 4% Na; A-RDNx, 20% fructose + 4% Na with afferent renal denervation; T-RDNx, 20% fructose + 4% Na with total renal denervation. Scale is 500 µm for 4× and 100 µm for 20× magnification.

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  Figure 3. Daily 24-h average systolic, diastolic, and mean blood pressures and heart rate in male (A–C,G) and female (D–F,H) rat groups. SBP, systolic blood pressure; DBP, diastolic blood pressure; MAP, mean blood pressure; HR, heart rate; bpm, beats per minute. Symbols apply to all graphs; group names as in Figure 2. Values are mean ± SE; n values are as in Table 1. * p < 0.01 A-RDNx vs. G-Sham, # p < 0.02 F-Sham vs. G-Sham, † p < 0.05 F-Sham vs. G-Sham.

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  Figure 4. Average strain measurements for male (A–E) and female (F–J) groups by speckle-tracking echocardiography. Global longitudinal strain (GLS), GLS rate, fractional shortening, global circumferential strain (GCS and end systolic volume (ESV). Symbols apply to all graphs; group names as in Figure 2. Values are mean ± SE. * p < 0.05, ** p < 0.01.

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  Figure 5. Histologic evaluation of cardiac fibrosis. Representative picrosirius red staining of left ventricle tissues in the four groups of male (A) and female (C) rats. Quantification of fibrotic area is shown in males (B) and females (D), respectively. Scale bar is 50 µm. Group names as in Figure 2. Values are mean ± SE. * p < 0.05, # p < 0.01.

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  Figure 6. Aortic pulse wave velocity (PWV) in male (A) and female (B) rats. Group names as in Figure 2. Values are mean ± SE. * p < 0.05, # p < 0.01.

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  Figure 7. Ex vivo myography of thoracic aortic rings in male (A–C) and female (D–F) rats in response to phenylephrine (PE), acetylcholine (ACh), or sodium nitroprusside (SNP). Values are mean ± SE; n = 6 for all groups. * p < 0.0001 from baseline, † p < 0.006 vs. F-Sham, # p < 0.025 vs. G-Sham as shown by two-way ANOVA.

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  Figure 8. Aortic rings stained for superoxide with red mitochondrial superoxide indicator (MitoSOXTM), 4′6 diamidino-2-phenylindole (DAPI), and overlay from male (A) or female (C) rats in the four groups as indicated in Figure 2. Quantitation of immunofluorescence is shown in males (B) and females (D), respectively. Scale bar is 1000 µm. Values are mean ± SE. * p < 0.05.

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  Figure 9. Periodic acid Schiff (PAS) staining of renal tissue depicting representative glomeruli from the four groups of male (A) and female (D) rats. Quantitative morphometric assessment for males (B,C) and females (E,F) shows percentage of glomeruli with mesangial hypercellularity: normal <4, mild 4–5, moderate 6–7 mesangial cells/area. No severe hypercellularity was noted. Number of observations indicate the number of kidneys evaluated; 40 glomeruli were evaluated blindly per kidney. * p < 0.05, ** p < 0.01, *** p < 0.001.

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  Figure 10. Renal function assessed as plasma creatinine (PCr; (A,B)) and urinary albumin to creatinine ratio (Alb/Cr; (C,D)) in the four groups of rats as in Figure 2. Values are mean ± SE. * p < 0.05, ** p < 0.01.