Scoparone alleviates Ang II‐induced pathological myocardial hypertrophy in mice by inhibiting oxidative stress

Figures

• 

  FIGURE 1. Scoparone inhibits Ang II infusion‐induced pathological cardiac hypertrophy in vivo. A, Heart weight /bodyweight ratio (HW/BW), heart weight/tibia length ratio (HW/TL) in mice from the four groups (n = 7 or 8 per group). B, Representative photographs of wheat germ agglutinin (WGA) stained sections and data of cardiomyocytes cross‐sectional area (CSA, n = 4).C, Echocardiographic parameters, including FS (fractional shortening), LVPWd (left ventricular posterior wall thickness end diastole), LVIDd (left ventricular internal diameter end diastole), LVIDs (left ventricular internal diameter end systole) and IVSd (interventricular septum thickness end diastole) were evaluated and calculated (n = 7 or 8). D, Real‐time PCR analysis of ANP (atrial natriuretic peptide), BNP (brain natriuretic peptide) and β‐myosin heavy chain (β‐MHC) mRNA level (n = 4).*P < 0.05 versus control group; # P < 0.05 versus Ang II group

• 

  FIGURE 2. Scoparone inhibits Ang II infusion‐induced cardiac fibrosis in vivo. A, Masson's stain of left ventricle in mice from the four groups (n = 4). B, Immunofluorescence of α‐smooth muscle actin (α‐SMA, green) and vimentin (VIM, red, marker of fibroblasts), percentage of α‐SMA‐positive cardiac fibroblasts(CFs) were calculated (n = 4). C, level of fibrosis related proteins (n = 3).*P < 0.05 versus control group; # P < 0.05 versus Ang II group

• 

  FIGURE 3. Scoparone inhibits Ang II infusion‐induced oxidative stress of hearts in vivo. A, 4‐HNE immunohistochemical staining of left ventricular (LV) (n = 4). B, MDA level of hearts, results are expressed as nmol/mg of protein (n = 7‐8). C, measure of RAC1‐GTP, total RAC1, NOX2 and NOX4 in hearts with Western blot (n = 3). D, Rac1‐GTP immunofluorescence staining in cardiomyocytes (CMs, α‐actinin positive) in vivo (n = 4). E, Rac1‐GTP immunofluorescence staining in cardiac fibroblasts (CFs, vimentin positive) in vivo (n = 4).*P < 0.05 vs control group; # P < 0.05 vs Ang II group

• 

  FIGURE 4. Scoparone inhibits Ang II‐induced cardiomyocytes oxidative stress by suppressing Rac1. NRCMs were treated with Ang II (1 μmol/L), scoparone (SCO, 50 μmol/L), control or RAC1 overexpression adenovirus (10 moi) for 48 h. A, Western blot of RAC1‐GTP, t‐RAC1, NOX2 and NOX4 (n = 3); B, Immunofluorescent staining of RAC1‐GTP (n = 3); C.ROS assay (n = 3). *P < 0.05 vs control group; # P < 0.05 vs Ang II group; † P < 0.05 vs Ang II + SCO group

• 

  FIGURE 5. Scoparone inhibits Ang II‐induced cardiomyocyte hypertrophy by suppressing Rac1 mediated oxidative stress. NRCMs were treated with Ang II (0.1 μmol/L), scoparone (SCO, 50 μmol/L), control or RAC1 overexpression adenovirus (10 moi) for 48 h. A, 3H‐leucine incorporation assay (n = 4). B, Measure of cardiomyocytes surface area with immunofluorescence staining of cardiac troponin I (CTNI) (n = 8). C, Real‐time PCR analysis of ANP, BNP and β‐myosin heavy chain (β‐ MHC) mRNA level (n = 3).*P < 0.05; # P < 0.05 vs Ang II group; † P < 0.05 vs Ang II + SCO group

• 

  FIGURE 6. Scoparone inhibits Ang II‐induced cardiac fibroblasts oxidative stress by suppressing Rac1. NRCFs were treated with Ang II (1 μmol/L), scoparone (SCO, 50 μmol/L), control or RAC1 overexpression adenovirus (10 moi) for 48 h. A, Western blot of RAC1‐GTP, t‐RAC1, NOX2 and NOX4 (n = 3); B, immunofluorescent staining of p‐RAC1 (n = 3); C, ROS assay (n = 3). *P < 0.05 vs control group; # P < 0.05 vs Ang II group; † P < 0.05 vs Ang II + SCO group

• 

  FIGURE 7. Scoparone inhibits Ang II‐induced cardiac fibroblasts differentiation and collagen synthesis by suppressing Rac1. NRCFs were treated with Ang II (1 μmol/L), scoparone (SCO, 50 μmol/L), control or RAC1 overexpression adenovirus (10 moi) for 48 h. A, Immunofluorescent staining of α‐SMA (n = 3). B, Western blot of α‐SMA, CTGF, collagen I and collagen III (n = 3). *P < 0.05 vs control group; # P < 0.05 vs Ang II group; † P < 0.05 vs Ang II + SCO group