Cardiac remodeling in high PDCD5 over-expressing line.

<p>(A), Cardiac specific high over-expression of hPDCD5 results in enlarged hearts (3 months old). (B), HW∶BW ratio showing significant increase in cardiac mass in high over-expressing line compared to the WT control mice (n = 6 for TG, n = 6 for WT). (C), Measurement of two-dimensional cardiomyocyte cross-sectional area showing significantly enlarged cells in high over-expressing line compared with WT control mice (n = 4 for TG, n = 4 for WT). *<i>P</i><0.05, WT vs. TG. (D), Histological analysis with H&E staining on heart sections. Bar, 50 µm. (E), Masson's trichrome staining showing collagen depositon in left ventricle of high over-expressing line. Bar, 50 µm. Expression of ANF (F), BNP (G), SAA (H), αMHC (I), βMHC (J) and SERCA (K) was determined by quantitative Real-Time RT-PCR analysis in cDNA samples derived from heart of high over-expressing line and WT control mice (n = 4 for TG, n = 4 for WT). Expression levels were normalized to GAPDH. Experiments were performed twice in triplicate with similar results. *<i>P</i><0.05, WT vs. TG.</p

Susceptibility of low over-expressing line to Ang II-induced cardiac hypertrophy.

<p>(A), H&E staining on heart sections. Bar, 40 µm. (B), Measurement of two-dimensional cardiomyocyte cross-sectional area. (C), LV to BW ratio is significantly increased in Ang II- treated transgenic mice as compared to WT control mice receiving identical treatment. (D), In transgenic mice treated with Ang II, FS% is not significantly altered as compared to WT control mice. (E), Representative images of LC3 processing in Ang II-treated transgenic mice and WT control mice. (F) Densitometric analysis of LC3 immunoblots. *<i>P</i><0.05, WT vs. TG (n = 4–5 for each group).</p

The effect of hPDCD5 over-expression on the survival of mice.

<p>(A), Autopsy of transgenic founder 41 showing dramatically enlarged heart. (B), Kaplan-Meier Survival Curves of high over-expressing line (red line) and WT littermate controls (black line). (n = 50 for WT, n = 65 for TG).</p

CR-SPRC reduced infarct size of left ventricle.

<p>(A) Representative photograph of infarct size which was determined by 1% triphenyltetrazolium chloride (TTC) staining. (B) Statistical analysis of infarct size. Data were presented as means ± standard deviations (n = 5–8). ^#<i>P</i><0.01 versus sham, ^*<i>P</i><0.01 versus HF, ^&<i>P</i><0.01 versus CR-SPRC.</p

Up-regulation of PDCD5 in Ang II-induced cardiac hypertrophy.

<p>(A), HW∶BW ratio showing significant increase in cardiac mass in mice treated with Ang II for 2 weeks compared to sham control mice. (B) Expression of ANF was determined by quantitative Real-Time RT-PCR analysis in cDNA samples derived from hearts of mice treated with Ang II and sham control. (C), Expression of βMHC was determined by quantitative Real-Time RT-PCR analysis in cDNA samples derived from hearts of mice treated with Ang II and sham control. (D), Representative western blot of PDCD5 and internal control actin proteins in heart extracts from 8–week-old male mice with sham or Ang II treatment. (E), Quantitative analysis revealed that PDCD5 levels were up-regulated in hearts from mice treated with Ang II (n = 4) as compared to sham control mice (n = 4). *<i>P<0.05</i>, sham vs. Ang II treatment.</p

Oligos for Real-Time RT-PCR experiments.

<p>Oligos for Real-Time RT-PCR experiments.</p

Assessment of autophagy in high over-expressing line.

<p>(A), Representative images of Immunostaining of LC3 on paraffin section of hearts from high over-expressing line and WT control mice. (B), Quantification of LC3 positive dots (For each group, 4 mice were studied). (C), Representative western blot of autophagy-related proteins LC3, Beclin 1, LAMP-1 and cathepsin D in heart extracts obtained from 3-month-old high over-expressing line and WT control mice. (D), Densitometric analysis of LC3 immunoblots. (E), Representative western blot of time course analysis of LC3 processing in heart extracts obtained from high over-expressing line and WT control mice. (F), Representative western blot of acetylated p53 and DRAM.</p

Echocardiographic analysis of cardiac function of high over-expressing line.

<p>(A), Representative M-mode echocardiography images in 3-month-old mice. (B)–(C), LV end-diastolic diameter (LVID;d) and LV end-systolic diameter (LVID;s) were significantly increased in high over-expressing line compared to WT control mice. (D), Left ventricular posterior wall (LVPW) was significantly decreased in high over-expressing line compared to WT control mice. (E)–(F), % fractional shortening and ejection fraction were significantly diminished in high over-expressing line compared to WT control mice. *<i>P</i><0.05, WT vs. TG (n = 5 for TG, n = 5 for WT, 3 months old).</p

Cardioprotective Effects of a Novel Hydrogen Sulfide Agent–Controlled Release Formulation of S-Propargyl-Cysteine on Heart Failure Rats and Molecular Mechanisms

<div><p>Objective</p><p>Heart failure (HF) is one of the most serious diseases worldwide. S-propargyl-cysteine (SPRC), a novel modulator of endogenous hydrogen sulfide, is proved to be able to protect against acute myocardial ischemia. In order to produce more stable and sustainable hydrogen sulfide, we used controlled release formulation of SPRC (CR-SPRC) to elucidate possible cardioprotective effects on HF rats and investigate involved mechanisms on apoptosis and oxidation.</p><p>Methods</p><p>Left coronary artery was occluded to induce HF model of rat. The survival rats were randomly divided into 7 groups after 24 hours and treated with drugs for 6 weeks. Echocardiographic indexes were recorded to determine cardiac function. TTC staining was performed to determine infarct size. Plasmatic level of hydrogen sulfide was detected by modified sulfide electrode. Activity of enzyme and expression of protein were determined by colorimetry and Western blot, respectively.</p><p>Results</p><p>The cardioprotective effects of CR-SPRC on HF rats were confirmed by significant reduction of infarct size and improvement of cardiac function, with better effects compared to normal SPRC. CR-SPRC modulated antioxidant defenses by preserving levels of GSH, CAT and SOD and reducing CK leakage. In addition, CR-SPRC elevated ratio of Bcl-2/Bax and inhibited activity of caspases to protect against myocardial apoptosis. The cardioprotective effects of CR-SPRC were mediated by hydrogen sulfide.</p><p>Conclusions</p><p>All experiment data indicated cardioprotective effects of CR-SPRC on HF rats. More importantly, CR-SPRC exerted better effects than normal SPRC in all respects, providing a new perspective on hydrogen sulfide-mediated drug therapy.</p></div

CR-SPRC ameliorated myocardial fibrosis of left ventricle.

<p>(A) Representative photograph of myocardial fibrosis which was determined by Masson's trichrome staining. (B) Representative high-magnification microphotograph of Masson-stained sections from border zone in indicated groups in (A). Magnification: ×400. (C) Statistical analysis of myocardial fibrosis. Data were presented as means ± standard deviations (n = 4–6). ^#<i>P</i><0.01 versus sham, ^*<i>P</i><0.01 versus HF.</p