Resistance exercise training and blueberry extract protect against cardiac and skeletal muscle remodeling and metabolism disruptions in experimental pulmonary arterial hypertension

Figures

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  Fig. 1. Schematic of experimental design adopted in this study. SH, SHB, EH, and EHB correspond to equivalent groups that received the same interventions but were euthanized on a fixed day (day 26) rather than being observed until failure. SH, sedentary hypertensive; SHB, sedentary hypertensive treated with blueberry; EH, exercised hypertensive; EHB, exercised hypertensive treated with blueberry; RT, resistance training; SHF, sedentary hypertensive until failure; SHBF, sedentary hypertensive treated with blueberry extract until failure; EHF, exercised hypertensive until failure; EHBF, exercised hypertensive treated with blueberry extract until failure; SC, sedentary control; MLT, maximum load test; PAH, pulmonary arterial hypertension; BL, blood lactate; Echo, echocardiography.

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  Fig. 2. Effects of resistance exercise training and blueberry extract on mortality rate, body weight and exercise tolerance assessed longitudinally until death. (A) Death proportion. (B) Death time. (C) Body weight throughout the experiment. (D) Relative maximum load in exercise tolerance tests. Data are means ± standard deviation of 8–10 rats in each group. The dotted lines indicate the dates of death of the last animals in each group. The red dashed rectangle highlights the region where significant changes in body weight were observed. One-way analysis of variance followed by the Tukey’s post hoc test was used for test differences between groups for body weight and relative carrying load. Kaplan-Meier curve followed by the Log-rank post hoc test for mortality proportion. The solid line (───) in panel A represents the median survival time (time after MCT injection when > 50% of the group died). MCT, monocrotaline; SC, sedentary control; SHF, sedentary hypertensive until failure; SHBF, sedentary hypertensive treated with blueberry extract until failure; EHF, exercised hypertensive until failure; EHBF, exercised hypertensive treated with blueberry extract until failure. aP < 0.05, SHF vs.…

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  Fig. 3. Effects of resistance exercise training and blueberry extract on pulmonary artery resistance, cardiac function, and RV single myocyte dimension and function assessed 23 days after monocrotaline injection. (A) Representative images of pulmonary artery flow. (B) Representative images of TAPSE. (C) Representative images of right and LV, and interventricular septum echocardiography. (D) Heat rate. (E) AT/ET ratio in the pulmonary artery. (F) TAPSE. (G) D-shaped LV. (H) Ejection fraction. (I) Fractional shortening. (J) Sarcomere length and width in isolated RV myocytes. (K) Time to 90% peak sarcomere shortening in isolated RV myocytes. (K) Time to 90% relaxation in isolated RV myocytes. Sectioned lines represent threshold values for dysfunction. White arrows indicate the flattening of the interventricular septum. Data are means ± standard deviation of 8 rats in each group. One-way analysis of variance followed by the Tukey’s post hoc test was used for test differences between groups for all variables and χ2 test to assess the differences in proportions between the groups. RV, right ventricle (or right ventricular); TAPSE, tricuspid annular plane systolic excursion; AT, acceleration…

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  Fig. 4. Effects of resistance exercise training and blueberry extract on the gene expression of metabolic function biomarkers in the RV and BB. (A) PGC1-α in the RV. (B) Pdk4 in the RV. (C) Cpt1b in the RV. (D) PGC1-α in the BB. (E) Pdk4 in the BB. (F) Cpt1b in the BB. Data are means ± standard deviation of 6–8 rats in each group. One-way analysis of variance followed by Tukey’s post hoc test was used for test differences between groups for all variables. RV, right ventricle (or right ventricular); BB, biceps brachii; PGC1-α, peroxisome proliferator-activated receptor gamma coactivator 1-alpha; Pdk4, pyruvate dehydrogenase kinase 4; Cpt1b, carnitine palmitoyltransferase 1B; SC, sedentary control; SH, sedentary hypertensive; SHB, sedentary hypertensive treated with blueberry; EH, exercised hypertensive; EHB, exercised hypertensive treated with blueberry. *P < 0.05 vs. SC; #P < 0.05 vs. SH; &P < 0.05 vs. SHB.

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  Fig. 5. Effects of resistance exercise training and blueberry extract on the oxidative stress biomarkers in the biceps brachii. (A) SOD. (B) CAT. (C) GST. (D) MDA. (E) NO. (F) PC. Data are means ± standard deviation of 6–7 rats in each group. One-way analysis of variance followed by Tukey’s post hoc test was used for test differences between groups for all variables. SOD, superoxide dismutase; CAT, catalase; GST, glutathione S-transferase; MDA, malondialdehyde; NO, nitric oxide; PC, protein carbonyl; SC, sedentary control; SH, sedentary hypertensive; SHB, sedentary hypertensive treated with blueberry; EH, exercised hypertensive; EHB, exercised hypertensive treated with blueberry. *P < 0.05 vs. SC; #P < 0.05 vs. SH.

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  Fig. 6. Effect of resistance exercise training and blueberry extract on the expression of proteins in the physiological hypertrophy pathway of the right ventricle. (A) PI3K. (B) Akt. (C) p-Akt. (D) mTOR. (E) p-mTOR. (F) 4E-BP1. (G) p-4E-BP1. (H) eIF4E. (I) p-eIF4E. (J) GSK3β. (K) p-GSK3β. (L) FOXO3. (M) p-FOXO3. (N) p-Bad. (O) B-cell lymphoma-extra-large (Bcl-xL). Data are means ± standard deviation of 6–8 rats in each group. One-way analysis of variance followed by Tukey’s post hoc test was used for test differences between groups for all variables. PI3K, phosphoinositide 3-kinase; Akt, protein kinase B; p-Akt, phospho-Akt; mTOR, mammalian target of rapamycin; p-mTOR, phospho-mTOR; 4E-BP1, eukaryotic translation initiation factor 4E-binding protein 1; p-4E-BP1, phospho-4E-BP1; eIF4E, eukaryotic translation initiation factor 4E; p-eIF4E, phospho-eIF4E; GSK3β, glycogen synthase kinase 3 beta; p-GSK3β, phospho-GSK3β; FOXO3, forkhead box O3; p-FOXO3, phospho-FOXO3; p-Bad, phosphorylated Bcl-2 antagonist of cell death; SC, sedentary control; SH, sedentary hypertensive; SHB, sedentary hypertensive treated with blueberry; EH, exercised hypertensive; EHB, exercised hypertensive treated with…

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  Fig. 7. Schematic representation of the main findings on the RT and blueberry extract protection against the progression of cardiac and skeletal muscle remodeling and metabolism disruptions in the MCT-induced PAH model. Upward arrows indicate an increase, while downward arrows indicate a reduction. Red arrows represent the effects of hypertension; green arrows indicate the exercise effects and blue arrows correspond to blueberry extract effects. RT, resistance exercise training; MCT, monocrotaline; PAH, pulmonary arterial hypertension; BW, body weight; 4E-BP1, eukaryotic translation initiation factor 4E-binding protein 1; Akt, protein kinase B; BB, biceps brachii; eIF4E, eukaryotic translation initiation factor 4E; FOXO3, forkhead box O3; GSK3β, glycogen synthase kinase 3 beta; IGF-1, insulin-like growth factor-1; IGF-1R, insulin-like growth factor-1 receptor; IRS-1, insulin receptor substrate 1; LV, left ventricle (or left ventricular); mTOR, mammalian target of rapamycin; P70S6K, ribosomal protein S6 kinase beta-1; PDK1, pyruvate dehydrogenase kinase 1; PI3K, phosphatidylinositol 3-kinase; PIP2, phosphatidylinositol 4,5-bisphosphate; PIP3, phosphatidylinositol (3–5)-triphosphate; RV,…