Distribution of Cardiac and Renal Corin and Proprotein Convertase Subtilisin/Kexin-6 in the Experimental Model of Cardio-Renal Syndrome of Various Severities

Figures

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  FIGURE 1. Pressure-volume loops in compensated and decompensated CHF and their sham controls. (A) Representative pressure-volume loops of rats with compensated CHF, decompensated CHF, and sham controls. (B) Heart rate, (C) stroke volume, (D) cardiac output, (E) ejection fraction, (F) end-systolic volume, (G) end-diastolic volume, (H) stroke work, (I) dP/dTmax, (J) dP/dTmin. Ejection fraction equals (end-diastolic volume – end-systolic volume)/end diastolic volume. Stroke work is calculated as the area under the pressure-volume curve (volume*pressure product). Maximum and minimum dP/dt signify the maximal systolic pressure generation as a parameter of left ventricular contractility and the minimal diastolic pressure loss as a parameter of left ventricular relaxation capacity, respectively. Values are means ± SEM.

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  FIGURE 2. Tissue fibrosis in ACF rats. Representative images of Masson’s trichrome staining of the cardiac ventricular and renal tissues, as well as renal cortical and medullar layers, illustrating the impact of ACF placement on the development of tissue remodeling, compared with sham operated rats. Images are presented at x0.6, x5, and x20 magnification.

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  FIGURE 3. Levels and expression of corin and PCSK6 in the cardiac tissue of compensated CHF, decompensated CHF, and sham controls. Representative western blot analysis of corin and PCSK6 in RA, LA, RV, and LV tissue lysates are shown in (A,B,K,L), respectively. (C–F,M–P) Represent western blot analysis relative quantification of corin and PCSK6 in the different heart chambers, where GAPDH was used as a loading control. Quantification of qPCR analysis for corin and PCSK6 mRNA normalized to GAPDH are depicted in (G–J,Q–T). Values are means ± SEM.

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  FIGURE 4. Corin/PCSK6 staining in the heart chambers of ACF and sham-operated rats. Representative images of immunofluorescence staining for corin (A) and PCSK6 (B) in the cardiac atrial and ventricular tissues of rats with compensated CHF, decompensated CHF, and sham-operated animals. Tissues were counterstained with DAPI. All images were obtained at x40 magnification.

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  FIGURE 5. Natriuretic peptide machinery system in the renal tissue of ACF and sham-operated rats. Representative western blot analysis (A) with antibodies against corin, PCSK6, ADAM10, and NEP and their quantification relative to GAPDH (B–E) in the kidney of ACF rats and their sham operated rats. (F–I) represents quantification of qPCR analysis of the mRNA of these enzymes relative to GAPDH. (J) Representative images of immunostaining of corin and PCSK6 in the kidney of the various animal groups, counterstained with DAPI. All images were obtained at x40 magnification. Values are presented as means ± SEM.

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  FIGURE 6. Abundance and expression of ADAM10 in heart chambers of ACF and sham-operated rats. Representative western blot analysis of ADAM10 (A,D,G,J) and its quantification relative to GAPDH (B,E,H,K) in RA, LA, RV, and LV, respectively, of ACF and sham-operated rats. (C,F,I,L) Represent quantification of qPCR analysis of ADAM10 mRNA relative to GAPDH in the heart chambers of the different animal groups. Values are presented as means ± SEM.

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  FIGURE 7. Abundance and expression of atrial and B-type natriuretic peptides in the cardiac and renal tissues of ACF and sham-operated rats. Representative western blot analysis of proANP (A,D,G) and proBNP (J,M,P) and their quantification relative to GAPDH (B,E,H,K,N,Q) in heart chambers and kidneys of ACF and sham-operated rats. (C,F,I,L,O,R) Represent quantification of qPCR analysis of proANP and proBNP mRNA relative to GAPDH in the same tissues. Values are presented as means ± SEM.