Characterization of biventricular alterations in myocardial (reverse) remodelling in aortic banding-induced chronic pressure overload

Figures

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  Figure 1. Effect of pressure overload on left ventricular hypertrophy, fibrosis, LV passive tension and extracellular matrix remodelling. (A) Cardiomyocytes area evaluated by histology; Western blot relative quantification of: (B) ratio of phosphorylated to total AKT; (C) ratio of phosphorylated to total mTOR; (D) ratio of phosphorylated to total glycogen synthase kinase 3 beta (GSK3β); (E) cardiomyocytes passive tension; (F) ratio of S26 phosphorylation to total Titin PEVK segment; (G) ratio of pPKCα to total PKCα phosphorylation; (H) myocardial fibrosis; (I) mRNA expression of procollagen type-I; (J) mRNA expression of procollagen type-III; (K) lysyl oxidase; (L) mRNA expression of Transforming growth factor beta, (M) Interleukin-6; (N) ratio of phospho to total Signal transducer and activator of transcription 3; (O) representative images of cardiomyocyte hypertrophy and myocardial fibrosis; (P) representative western blot lanes. For representative purposes the image was cropped and edited; (Q) representative membrane protein loading. n = 7 for each group. Values are mean ± SEM, Two-way ANOVA for passive tension, One-way ANOVA for the remaining data. Ba/Deb vs Sh: αp < 0.05; ααp < 0.01;…

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  Figure 2. Effect of pressure overload on LV cardiomyocytes and calcium handling. (A) Cardiomyocytes active tension; (B) myofilaments calcium sensitivity; (C) Hill-coefficient; (D) the rate of tension redevelopment (Ktr); protein quantification of: (E) sarcoplasmic/endoplasmic reticulum Ca2+ATPase 2a (SERCA2a); (F) ratio of SERCA2a to phospholamban; (G) sodium-calcium exchanger; (H) ratio of phosphorylated to total Ca2+/calmodulin-dependent protein kinase (CAMKii); mRNA expression of calcium-binding protein A1 (S100A1); (J) representative western blot lanes. For representative purposes the image was cropped and edited; (K) representative membrane protein loading. Values are mean ± SEM. One Way-ANOVA: Ba/Deb vs Sh: αp < 0.05; ααp < 0.01; αααp < 0.001; Deb vs Ba: χp < 0.05; χχp < 0.01, χχχp < 0.001.

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  Figure 3. Effect of pressure overload on RV hypertrophy and fibrosis. (A) Foxo transcription factor 1; (B) ratio of phosphorylated to total protein kinase B or AKT; (C) ratio of phosphorylated to total P38 mitogen-activated protein kinase; (D) ratio of phosphorylated to total mammalian target of rapamycin (mTOR); (E) ratio of phosphorylated to total glycogen synthase kinase 3 beta (GSK3β); (F) ratio of phosphorylated to total extracellular signal–regulated kinase (ERK); (G) Myostatin; (H) Muscle-specific RING finger protein-1 (MuRF1); (I) representative western blot lanes (for representative purposes the image was cropped and edited) representative membrane protein loading. n = 5 for each group. Values are mean ± SEM, One-way ANOVA. Ba/Deb vs Sh: αp < 0.05; ααp < 0.01; αααp < 0.001; Deb vs Ba: χp < 0.05; χχχp < 0.001.

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  Figure 4. Effect of pressure overload on RV cardiomyocytes and calcium handling. (A) cardiomyocytes passive tension; (B) cardiomyocytes active tension; (C) myofilaments calcium sensitivity; (D) Hill-coefficient; (E) the rate of tension redevelopment (Ktr); Protein quantification of: (F) sarcoplasmic/endoplasmic reticulum Ca2+ATPase 2a (SERCA2a); (G) ratio of phospholamban phosphorylated (p-PLB) to total (PLB); (H) ratio of SERCA2a to phospholamban; (I) sodium-calcium exchanger (NCX); (I) Junctophilin; (L) representative western blot lanes (for representative purposes the image was cropped and edited) representative membrane protein loading. n = 5 for each group. Values are mean ± SEM, One-way ANOVA. Ba/Deb vs Sh: αp < 0.05; ααp < 0.01; αααp < 0.001; Deb vs Ba: χp < 0.05; χχχp < 0.001.

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  Figure 5. Effect of pressure overload on lung arteries thickness. (A–C) Pulmonary arterial thickness (D) representative images of histological lungs. Correlations between arterial wall thickness and: (E) maximum rate of oxygen consumption (VO2max); (F) ratio between: (E), peak of pulse Doppler wave of early mitral flow velocity and E′, wave velocity of tissue Doppler at the lateral mitral annulus; (E/e′) and (G) Indexed left ventricle area (LAAind). n = 8 for each group. Values are mean ± SEM, One-way ANOVA Ba/Deb vs Sh: αp < 0.05; ααp < 0.01; αααp < 0.001; Deb vs Ba: χp < 0.05; χχχp < 0.001.

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  Figure 6. Experimental protocol design.