A: Remaining ventricular tissue part stained with H&E. B: ratios of heart fat/body weight (HW/BW) and still left ventricular excess weight/human body weight (LVW/BW). C: Agent of H&E staining of 1352608-82-2 cost coronary heart tissue D: Statistical information of myocyte floor area proven in H&E staining calculated and analyzed 
with LAS Software program (V4.three) (Leica, Germany). E: mRNA expression of ANP. F: mRNA expression of BNP. Data are presented as suggest .D. n = eight for A, B, C and D n = 4 for E and F. : P<0.05 is considered statistical significance.Fig 2. Effect of ASIV on Iso-induced NRVM cells. A: The cell morphology was visualized by immunofluorescence analysis. B: Cell surfaced area was measured and analyzed with LAS Software (V4.3) (Leica, Germany). C: mRNA expression of ANP. D: mRNA expression of BNP. E: Protein content. Data are presented as mean .D. n = 4. : P<0.05 is considered statistical significance.Fig 3. Effects of ASIV on Iso-induced changes in the energy biosynthesis and expression of ATP5D. A and E: ATP/AMP ratio in vivo or in vitro. B and F: FFA level in vivo or in vitro. C and G: Representative of western blot photograph of ATP5D in vivo or in vitro. D and H: Statistical data of protein expression of ATP5D. Data are presented as mean .D. n = 8 for A, B, C and D n = 4 for E, F, G and H. : P<0.05 is considered statistical significance.Fig 4. Effects of ASIV on p65 and PGC-1 protein expression measured by Western blot. A and B: p65 expression in cytoplasm and nuclear fraction of heart tissue. C and D: p65 expression in cytoplasm and nuclear fraction of NRVMs.B: E and F: PGC-1 expression in heart tissue and NRVMs. Data are presented as mean .D. n = 4. : P<0.05 is considered statistical significance.increased the protein expression of PGC-1 in NRVMs compared with the Iso alone (Fig. 5), suggesting the importance of NF-B activation in the myocytes hypertrophy and downregulation of PGC-1.We reported in the present study that ASIV improves the cardiac hemodynamics, downregulates mRNA expression of ANP and BNP, corrects the dysfunction of energy biosynthesis, up-regulates the protein expression of ATP5D and PGC-1, and inhibited the translocation of p65 into nuclear fraction from cytoplasm in rats and NRVMs treated with Iso. The findings confirm and expand the results reported previously[21], in which we found that ASIV protects against Iso-induced cardiac hypertrophy in terms of ratios of HW/BW and LHW/BW as well as ANP and BNP mRNA expression. In the present study, we additionally examined the cardiac hemodynamics and verified the protective role of ASIV in cardiac hypertrophy based on the improvement of hemodynamics. Cardiac hypertrophy is commonly viewed as a compensatory response, accompanied by neurohormonal activation, inflammatory responses and metabolic remodelling[1,2]. The energy metabolic reprogramming are characterized with mitochondrial dysfunction, shift of cardiac substrate utilization, and alterations in high-energy phosphate metabolism[17]. Clinical research revealed that ASIV had a variety of therapeutic effects on failure heart such as metabolism modulation, anti-inflammation, anti-oxidation, anti-thrombosis. The metabolism modulation of ASIV involved energy changes and ATPase activity[23,27]. It is known that ATP is required for myocardial pump function. The energy demands for the heart are immense. Normally, myocardial ATP is mainly generated from mitochondrial oxidation of fatty acids, which17600836 accounts for 600% of the total energy production[28]. Numerous experimental and clinical studies indicates that the hypertrophied heart is characterized by a shift in the utilization of substrate, from fatty acids oxidation towards glycolysis[29,30]. As the disease stage progressed, the presumably adaptive increase in glycolysis is not sufficient to meet ATP demand, following marked intracellular lipid accumulation and leading to lipotoxic cardiomyopathy[18].