Alda‑1, an aldehyde dehydrogenase‑2 agonist, improves long‑term survival in rats with chronic heart failure following myocardial infarction

Figures

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  Figure 1.. Long-term treatment with Alda-1 increases survival rate of CHF in post-MI rats. (A) ALDH2 activity in heart tissue from each group; 3 replicates of each sample, n=4/group. (B) ALDH2 protein expression levels were determined by western blot analysis using ALDH2 specific antibody; GAPDH was used as the loading control. (C) Kaplan-Meier survival curves and Mantel-Cox log rank test were used to examine the survival rates for rats in the Sham, MI and Alda-1 groups. Alda-1 treatment significantly improved 20-week survival rates; P=0.0297, Alda-1 vs. MI. Sham, n=17; MI, n=9; Alda-1, n=15. (D) BNP level in serum from each group; 3 replicates of each sample, n=4/group. ^P<0.01 vs. Sham, ^^P<0.001 vs. Sham, **P<0.001 vs. MI. ALDH2, alcohol dehydrogenase 2; BNP, B-type natriuretic protein; MI, myocardial infarction.

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  Figure 2.. Long-term treatment with Alda-1 attenuates ventricular remodeling and improves cardiac function. (A) Representative H&E stained cross-section images of heart (top); representative H&E staining micrograph images of left ventricles in non-infarcted area in each group (bottom); magnification, ×400. (B) Myocardial size is summarized and demonstrated; n=4/group. (C) HW/BW ratios in each group. (D) Echocardiography was performed 20 weeks following treatment. Representative images of 2D-guided M-mode echocardiographic of the left ventricle in each group are exhibited. (E) Quantitative analysis of LVIDd. (F) Quantitative analysis of LVIDs. (G) Quantitative analysis of LVEF. (H) Quantitative analysis of LVFS. Sham, n=17; MI, n=9; Alda-1, n=15; ^^P<0.001 vs. Sham, **P<0.001 vs. MI. H&E, hematoxylin and eosin; HW/BW, heart weight/body weight; LVEF, left ventricular ejection fraction; LVFS, left ventricular fractional shortening; LVIDd, left ventricular dimension at end diastole; LVIDs, left ventricular dimension at end systole.

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  Figure 3.. Long-term treatment with Alda-1 inhibits collagen formation in chronic heart failure post-MI rats. (A) Representative Mallory's trichrome staining micrographs demonstrated collagen fibers in non-infracted area of left ventricles; magnification, ×400. (B) Quantitative analysis of fibrotic area; Mallory-positive stained area (light blue color) was normalized to total myocardial area; n=4/group. (C) Representative images of collagen type I and type III protein expression in each group. (D) The ratio of collagen type I/typeIII were summarized; n=4/group. ^^P<0.001 vs. Sham, **P<0.001 vs. MI. MI, myocardial infarction.

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  Figure 4.. Long-term treatment with Alda-1 reduces myocyte apoptosis in chronic heart failure post-MI rats. (A) Representative images of cardiomyocyte apoptosis by Terminal deoxynucleotidyl-transferase-mediated dUTP nick-end labeling assay in each group; myocytes were detected with anti-MYH7 antibody (red) and nuclei were counterstained with DAPI (blue). (B) Quantification of apoptosis from (A), expressed as percentage of apoptotic cells out of the total number of cardiac cells; n=4/group. (C) Caspase-3 activity in heart tissue from each group; 3 replicates of each sample, n=4/group. (D) Protein expression levels of cleaved-caspase-3 were determined by western blot analysis; GAPDH was used as the loading control. ^^P<0.001 vs. Sham, **P<0.001 vs. MI. MI, myocardial infarction; MYH7, myosin-7.

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  Figure 5.. Long-term treatment with Alda-1 increases toxic aldehydes 4-HNE clearance. (A) Accumulation of 4-HNE adducts in the Sham group, MI group, Alda-1 group were measured by immunochemistry and (B) quantification of 4-HNE accumulation was calculated as the percentage of 4-HNE-positive stained cells; n=4/group; ^^P<0.001 vs. Sham, **P<0.001 vs. MI. (C) Western blot representative image of 4-HNE modified proteins. 4-HNE, 4-hydroxynonenal; MI, myocardial infarction.