Cholesterol-Lowering Gene Therapy Prevents Heart Failure with Preserved Ejection Fraction in Obese Type 2 Diabetic Mice

Figures

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  Figure 1. Schematic representation of the study design.

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  Figure 2. Quantification of murine LDLr expression in the liver. Bar graph (A) illustrating murine LDLr protein levels quantified by western blot in the liver of C57BL/6J LDLr−/− mice (n = 2), of wild-type C57BL/6J mice (n = 3), and of AAV8-LDLr C57BL/6 LDLr−/− mice (n = 3) 17 weeks after gene transfer with 3 x 1012 genome copies/kg of AAV8-LDLr. All protein levels were normalized to the ß-tubulin protein level. Image of western blot is shown in panel (B). The first two lanes correspond to C57BL/6J LDLr−/− mice, the next three lanes illustrate wild type C57BL/6 J mice, and the final three lanes correspond to AAV8-LDLr-treated C57BL/6 LDLr−/− mice.

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  Figure 3. Time course of body weight (A) and blood glucose levels (B) in C57BL/6J LDLr−/− mice fed a standard chow (SC) diet or a high-sucrose/high-fat (HSHF) diet. Week 0 in panels (A) and (B) corresponds to the age of 12 weeks, the start of the HSHF diet. All data represent mean ± SEM (n = 24 for SC diet groups; n = 36 for HSHF diet groups).

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  Figure 4. AAV8-LDLr gene transfer abrogates adipocyte hypertrophy induced by the HSHF diet. Adipocyte cross-sectional area (A) and adipocyte density (B) in female C57BL/6J LDLr−/− mice at 16 weeks after the start of the diet. Data are expressed as means ± SEM (n = 5). (C) This panel contains representative photomicrographs illustrating haematoxylin and eosin-stained adipocytes of the gonadal fat pad. The scale bar represents 50 µm.

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  Figure 5. Plasma insulin (A), free fatty acids (B), and adiponectin (C) levels at 16 weeks after the start of the diet. All data represent means ± SEM (n = 10).

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  Figure 6. Total cholesterol (A), non-HDL cholesterol (B), HDL cholesterol plasma levels (C), and plasma triglyceride levels (D) in female C57BL/6J LDLr−/− mice at 16 weeks after the start of the diet. All data are represented as means ± SEM (n = 10).

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  Figure 7. AAV8-LDLr gene transfer attenuates liver steatosis induced by the HSHF diet. Liver steatosis expressed as percentage of surface area in female C57BL/6J LDLr−/− mice at 16 weeks after the start of the diet is shown in panel (A). Data are expressed as means ± SEM (n = 5). Panel (B) contains representative photomicrographs illustrating hematoxylin- and eosin-stained liver tissue. The scale bar represents 50 µm.

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  Figure 8. Representative Sirius Red-stained heart cross-sections of SC diet and HSHF diet mice. The scale bar represents 1 mm.

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  Figure 9. Immunohistochemical and histochemical analysis of the myocardium of SC diet and HSHF diet mice. Representative photomicrographs show laminin-stained cardiomyocytes, CD31-positive capillaries, Sirius Red-stained collagen, and 3-nitrotyrosin. The scale bar represents 50 µm.

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  Figure 10. Lactate pre-exercise (A), distance covered (B), and lactate post-exercise (C) in C57BL/6 LDLr−/− mice fed the SC diet or the HSHF diet. All data represent means ± SEM (n = 11 for SC diet mice, n = 15 for HSHF diet mice).

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  Figure 11. Plasma TNF-α concentration in C57BL/6J LDLr−/− mice fed the SC diet or the HSHF diet and in C57BL/6J mice fed the SC diet or the HSHF diet. All data represent means ± SEM (n = 10).