Preexisting heart failure with reduced ejection fraction attenuates renal fibrosis after ischemia reperfusion via sympathetic activation

Figures

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  Figure 1. Experimental schematic.

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  Figure 2. Characteristics of TAC mice. This Figure shows (A–E) the echocardiogram result, (F) the weight of heart measured as heart/tibia ratio, and (G) the area of heart fibrosis. The echocardiogram revealed that TAC mice had reduced fractional shorting (A), larger left ventricular end-diastolic and end-systolic diameter (LVDd, LVDs) and thickened interventricular septum (IVS) (B–D). Significant differences were observed in heart weight and fibrosis between the TAC mice and the non-TAC mice (F,G). *p < 0.05. Gray bar and open triangle, non-TAC mice; black bar and open circle, TAC mice. All graphs were created using JMP 14.0 software (SAS Institute Inc., Cary, NC).

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  Figure 3. TAC had no effect on the acute phase after renal IR (Experiment 1). (A) Systolic blood pressure before renal IR, (B) kidney weight per body weight (BW) 24 h after renal IR, and (C) BUN before and 24 h after renal IR were similar between the non-TAC (n = 5) and the TAC groups (n = 5). Gray bar, non-TAC mice; black bar, TAC mice. (D) Representative image of renal pathology at 24 h after renal IR in PAS staining. (Bar = 100 μm). All graphs were created using JMP 14.0 software (SAS Institute Inc., Cary, NC).

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  Figure 4. Renal interstitial fibrosis development after IR (Experiment 2). (A) Representative images of renal pathology with Masson Trichrome staining 14 days after IR. (B) Quantitative analysis of renal fibrotic areas (n = 4–5 per group). (C) Representative images of α-SMA staining 14 days after IR. (D) Quantitative analysis of α-SMA positive area (n = 4–5 per group). (E) mRNA levels of TGF-β is shown (n = 4–5 per group). *p < 0.05. Open triangle, non-TAC mice; filled circle, TAC mice. All graphs were created using JMP 14.0 software (SAS Institute Inc., Cary, NC).

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  Figure 5. Macrophage infiltration and MCP1 mRNA expression after IR (Experiment 2). (A) Representative images of immunohistochemistry for F4/80 before, at 1, 7 and 14 days after IR (Bar = 25 μm). (B) Number of F4/80 positive cells in high power field (× 400) and (C) MCP1 mRNA expression (n = 4–5 per group). *p < 0.05. Gray bar and open triangle, non-TAC mice; black bar and open circle, TAC mice. All graphs were created using JMP 14.0 software (SAS Institute Inc., Cary, NC).

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  Figure 6. Relative transcript level of genes related with anti-fibrotic pathway (Experiment 2). mRNA levels of (A) HIF1, (B) HO-1, (C) CSF2, (D) BMP7 and (E) ALK-3 in the kidneys of non-TAC and TAC mice are shown (n = 4–5 per group) after IR. Gray bar and open triangle, non-TAC mice; black bar and filled circle, TAC mice. All graphs were created using JMP 14.0 software (SAS Institute Inc., Cary, NC).

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  Figure 7. Renal sympathetic denervation worsened renal fibrosis after IR in TAC mice (Experiment 3). (A) Fractional Norepinephrine (NoE) turnover rate (%/h), (B) NoE concentration in kidney (ng/g kidney), (C) Quantified renal fibrotic areas and (D) α-SMA positive area, and (E) mRNA level of TGF-β are shown (n = 5–7 per group). #p < 0.05. In (B), open triangle represents non-TAC mice without RSDN and IR; gray triangle non-TAC + IR mice without RSDN; filled triangle non-TAC + IR with RSDN; open circle, TAC mice without IR and RSDN; gray circle, TAC + IR mice without RSDN; filled circle, TAC + IR + RSDN mice. In (C–E), open triangle represents non-TAC + IR mice without RSDN; filled triangle non-TAC + IR with RSDN; open circle, TAC + IR mice without RSDN; filled circle, TAC + IR + RSDN mice. All graphs were created using JMP 14.0 software (SAS Institute Inc., Cary, NC).