Perindopril Improves Cardiac Function by Enhancing the Expression of SIRT3 and PGC-1α in a Rat Model of Isoproterenol-Induced Cardiomyopathy

Figure 1. Perindopril (PER) enhances heart function in isoproterenol (ISO)-induced cardiomyopathy model rats. (A) Changes in body weights for control rats and ISO-induced cardiomyopathy rats treated with out without PER (n = 10). (B) Changes in rat heart weight index (HWI) for different groups (n = 10). (C) Ang II levels in sera were measured by ELISA assay (n = 6). (D) B-type natriuretic peptide (BNP) levels in sera were measured by ELISA assay (n = 6). (E) Histopathological changes in ventricular myocardial tissues observed by H&E staining (×200 magnification) (n = 5). (F) Masson’s Trichrome Staining of cardiac tissues (×200 magnification). Blue color represents fibrosis (n = 5). (G) The Collagen Volume Fraction (CVF) of myocardial tissues was calculated as quantification (n = 5). (H) Representative M-mode echocardiograms for ISO, ISO + PER and control groups (n = 6). (I) Bar charts showing LVEDD, LVESD, LVEF%, and LVFS%. Data are represented as mean ± SD, #P < 0.05, ##P < 0.01 compared with ISO group, *P < 0.05, **P < 0.01 compared with control group, ▲▲P < 0.01 compared with Week 2, LVEDD: Left ventricular end diastolic diameter; LVESD: Left ventricular end systolic diameter; LVEF%:…

Figure 2. Perindopril (PER) attenuates myocardial mitochondrial function and oxidative stress in isoproterenol (ISO)-induced cardiomyopathy rats. (A) Flow cytometry was performed to analyze the total reactive oxygen species (ROS) levels in podocytes based on DCHF-DA staining (n = 3). (B) Quantitative analysis of ROS (n = 3). (C) Myocardial tissues were homogenized, and then GSH-px, Mn-SOD, and malondialdehyde (MDA) levels were assessed (n = 6). (D) ATP content in myocardial tissues was assessed (n = 6). (E) Cardiac ultrastructure of myocardium examined by electron microscope, magnification 10,000× and 30,000× (n=3). (F) Mitochondria were isolated from myocardial tissues for detection of complexes I, II, III, IV activities (n = 6). Data are represented as means ± SD, ##P < 0.01 compared with the ISO group, and **P < 0.01 compared with the control group.

Figure 3. Perindopril (PER) attenuates myocardial apoptosis in isoproterenol (ISO)-induced cardiomyopathy rats. (A) Western blotting of Cyto Cyt C in cytoplasmic lysates, Mito CytC in mitochondrial lysates, and Cleaved Caspase-3 and Pro-Caspase-3 in total cell lysates (n = 3). (B) Flow cytometry was employed to analyze the mitochondrial membrane potential (MMP) of myocardial cells based on JC-1 staining (n = 3). (C) The MMP was quantified as the red-to-green fluorescence ratio. (D) Representative DNA fragmentation assays measured by DAPI and TUNEL staining. (E) The apoptosis rate was computed as the positive cell percentage. Statistics were calculated from 5 views from every group (n = 5). Data are represented as means ± SD, ##P < 0.01 compared with the ISO group, **P < 0.01 compared with the control group.

Figure 4. Perindopril (PER) restores the protein and mRNA levels of peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1α), Sirtuin 1 (SIRT1), mitochondrial transcription factor A (TFAM), and nuclear respiratory factor 1 (NRF1) in myocardial tissues from isoproterenol (ISO)-induced cardiomyopathy rats (n = 3). (A) Real-time PCR was carried out to analyze the mRNA levels of PGC1α, SIRT1, TFAM, and NRF1 within myocardial tissues (n = 3). (B) Western blotting of PGC1α, SIRT1, TFAM, and NRF1 within myocardial tissues (n = 3). Data are represented as means ± SD, #P < 0.05, ##P < 0.01 compared with the ISO group, **P < 0.01 compared with the control group.