Effects of Adiponectin on Diastolic Function in Mice Underwent Transverse Aorta Constriction

Figures

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  Fig. 1. Evaluation of cardiac function, structure, and morphology in three groups of mice. a M-mode echocardiography showed increased left ventricular posterior wall thickness (LVPW) in TAC mice while APN-treated mice demonstrated reduced LVPW. b Mitral inflow pattern and c mitral annular velocity of TAC group revealed progressive diastolic dysfunction while APN-treated mice showed recovery of diastolic dysfunction, n = 10 per group. d Representative global heart photographs. e Hematoxylin and eosin-stained LV transverse sections (original magnification × 400; sale bar, 20 μm). f Representative microscopic images of Masson’s trichrome staining (original magnification × 400; sale bar, 75 μm). g Quantitative analysis of heart weight to body weight ratio. h Quantitative analysis of heart weight to tibia length ratio. i Quantitative analysis of cross-sectional area (CSA). j Quantification of myocardial fibrotic area (%). k, l Expressions of hypertrophy-associated genes. GAPDH served as the internal control. ANF, natriuretic peptide A. BNP, natriuretic peptide B. Values represent the mean ± SEM, n = 6 per group. **P < 0.01; *P < 0.05

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  Fig. 2. APN improved active relaxation of single myocytes in TAC mice. a Resting cell length. b Peak shortening. c Maximal velocity of shortening (+dL/dt). d Maximal velocity of re-lengthening (−dL/dt). e Time-to-50% re-lengthening. f Time-to-90% re-lengthening. Values represent the mean ± SEM, n = 45–50 cells from 5 mice per group. **P < 0.01; *P < 0.05

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  Fig. 3. Titin isoform shift and extent of matrix extracellular reorganization in the left ventricular myocardium from three groups of mice. a Representative images of titin isoforms in mice LV from indicated groups. b Quantitative analysis of titin isoforms. c–e Representative blots of collagen type I, collagen type III, and Lox. f–i Quantitative analysis of collagen type I, collagen type III, collagen type I to III ratio, and Lox. Values represent the mean ± SEM, n = 6 per group. **P < 0.01; *P < 0.05

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  Fig. 4. APN abated gene expressions of NADPH oxidase subunits in TAC mice. a, b Representative protein blots of MnSOD and GAPDH in mice ventricular myocardium. Values represent the mean ± SEM, n = 6 per group. **P < 0.01. c–g Relative mRNA expressions of p22phox, gp91phox, p47phox, p67phox, and Rac1. Values represent the mean ± SEM, n = 6 per group. **P < 0.01; *P < 0.05

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  Fig. 5. AMPK inhibition abrogated effects of APN in regulating titin isoform transformation. a Representative protein expressions of phosphorylated AMPK, total AMPK in mice ventricular myocardium. b Quantitative analysis. Values represent the mean ± SEM, n = 6 per group. *P < 0.05. c Representative protein expressions of phosphorylated AMPK, total AMPK in cardiomyocytes. d Quantitative analysis. Values represent the mean ± SEM. Experiments were repeated five times with cells obtained from three independent cultures. *P < 0.05. e–g Cells were under stimulation of compound C at the concentration of 2, 4, and 6 μM, respectively. *P < 0.05; **P < 0.01. h Representative images of titin isoforms in NRCMs treated with AngII, in the presence or absence of APN or Cpc. i Quantitative analysis of titin isoforms. *P < 0.05. Values represent the mean ± SEM. Experiments were repeated five times with cells obtained from three independent cultures

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  Fig. 6. AMPK inhibition abrogated the anti-hypertrophic effect of APN. a Representative images of NRCMs under indicated stimulations. b Quantitative analysis of NRCMs size (original magnification, × 400). c, d Hypertrophic gene expressions of ANF and BNP. Values represent the mean ± SEM. Experiments were repeated five times with cells obtained from three independent cultures. *P < 0.05; **P < 0.01