Limited synergy of obesity and hypertension, prevalent risk factors in onset and progression of heart failure with preserved ejection fraction

Figures

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  Figure 1. Workflow and model characteristics. (A) Graphical workflow, showing that male lean and obese ZSF1 rats were studied from 12 to 26 weeks of age. Both lean and obese rats were treated with DS or placebo from 19 to 26 weeks of age to trigger severe hypertension. Echocardiographic, metabolic and hemodynamic analyses were done every 2 weeks, till sacrifice at the age of 26 weeks. Longitudinal plots of (B) systolic blood pressure (mm Hg), (C) ejection fraction (%) and (D) E/e’ ratio. n = 6‐7, * obese vs lean, § obese + DS vs lean + DS, ‡ lean + DS vs lean, † obese + DS vs obese P < .05. (E) Principal component analysis of expression profiles in cardiac tissue. Symbols denoting subgroups panels B‐E are shown in panel B. (F‐I) Graphic display of differential gene expression in which log2FC is plotted against the mean of normalized counts. Red dots represent the differentially expressed genes (FDR < 0.1), grey dots represent non‐differentially expressed genes

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  Figure 2. Pathway analysis and mitochondrial gene expression (A) Graphical visualization of 3 most affected pathways according to Ingenuity Pathway Analysis (orange and blue represent pathway activation and repression, respectively, grey represents no direction available). (B) Schematic presentation of RNAseq data for differentially expressed genes involved in fatty acid oxidation, (C) KLF15‐induced transcription (D) and oxidative phosphorylation. Shown is a colour‐based representation of the reads per kilobase million, relative to the average expression in the lean group, for each individual rat (orange and blue represent increased and decreased expression, relatively). (E) QPCR analysis of oxidative stress marker CAT. n = 5‐7, * P < .05. (F) Analysis of urinary TBARS excretion as marker of systemic oxidative stress. n = 6‐7, * P < .05

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  Figure 3. Cardiomyocyte hypertrophy and foetal gene expression (A) Typical examples and (B) quantification of histological staining for cardiac hypertrophy (Gomori). n = 4‐6, * P < .05. C, QPCR analysis of cardiac transcription of hypertrophy markers MYH6, MYH7 and NPPA, (D) regulators of cytoplasmic Ca2+ levels SERCA2A, PLN and ATP1A2 and (E) transcriptional target of Ca2+‐induced NFAT activation RCAN1. n = 5‐7, * P < .05

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  Figure 4. Capillary density. (A) Typical examples and (B) quantification of LV microvasculature (Lectin). n = 4‐6, * P < .05. (C) QPCR analysis of cardiac transcription of factors involved in vascular remodelling ANGPT1, ANGPT2 and VEGFA. n = 5‐7, * P < .05

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  Figure 5. Inflammatory cells. (A) Typical examples of immunological staining for LV T cell (CD3) and (B) macrophage (CD68) presence. Positively labelled cells are depicted with an arrowhead. (C) Quantification of immunological staining for LV T cell (CD3) and (D) macrophage (CD68) presence per image field. n = 4‐7, * P < .05. E, Magnified visualization of immunological CD68 labelling in DS‐treated obese rats, illustrative for the observed DS‐induced increase in apoptosis seen in regions with high cellular density

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  Figure 6. Fibrosis and TGFβ signalling. (A) Typical examples and (B) quantification of histological staining for cardiac collagen deposition (Picro Sirius Red). n = 6‐7, * P < .05. (C) QPCR analysis of cardiac transcription of extracellular matrix components COL1A1, COL3A1 and FN1, (D) activators of TGFβ signalling TGFβ1 and TGFβ2 and (E) transcriptional targets of TGFβ signalling POSTN and SPP1. n = 5‐7, * P < .05