| In recent years, the cardiac energy metabolism obstacle in the occurrence and development of heart failure gets more and more attention. In cells, mitochondria as important organelles, are not only the main organelles that generate energy by oxidative phosphorylation, playing irreplaceable role in energy metabolism, but also involved in the cell oxygen free radical production, programmed cell death, signal transduction, various ions transshipment across cell membrane and the steady state of electrolyte. In the whole life of activity cells, mitochondria acts as a critical role. Therefore, regular pattern of mitochondrial structure change and cardiac energy metabolism obstacle in the process of chronic cardiac reconstruction is worth studying. Based on the fact that improving myocardial reconstruction treatment can delay heart failure progress, we assume that the treatment will protect mitochondrial function. Mitochondria are organelles producing energy. So improve the myocardial cell reconstruction will benefit mitochondrial energy metabolization. The traditional Chinese medicine of Benefiting Vital Energy, Activating Blood Circulation and Warming Yang are able to restrain myocardial reconstruction, and may also be benefit mitochondrial structure and cardiac energy metabolism.Objective:1. Primary cultures of neonatal rat myocardial cells were performed to explore pathological problems about mitochondria changes in the process of myocardial hypertrophy.2. Explore the influences of traditional Chinese drugs which benefit vital energy, activate blood circulation and warm yang on mitochondria and energy metabolism in the process of restraining myocardial cell hypertrophy caused by AngⅡ.Methods:1. Neonatal myocardial cells were dispersed and cultured with angiotensinⅡ(AngⅡ). We detected the total protein content with BCA method and measured cell diameter with inverted microscope, to show the situation of cardiomyocyte hypertrophy.2. We detected mitochondrial membrane potential (A^m) with fluorescence microscope and mitochondrial single amine oxidase (MAO) activity with spectrophotometer and mitochondrial cytochrome oxidase (COX) activity and the damage percentage of mitochondrial outer membrane with microplate reader and protein expression of mitochondrial outer membrane Bcl-2, inner membrane ANT with western blot, to show the damage of mitochondrial construction and function in hypertrophic cardiomyocyte. 3. We detected the content of ATP,ADP,AMP with high performance liquid chromatography and calculated the total adenosine acid content and energy charge, to show the situation of energy metabolism in hypertrophic cardiomyocyte.4. Valsartan as positive control, cells also were treated with ligustrazine,radix astragali injection and shenfu injection and then the pharmacological effects on mitochondrial structure, function and energy metabolism in the myocardial cells treated with AngⅡwere observed. 5. Observe the influences of energy metabolism drug(trimetazidine) on mitochondrial structure,function and energy metabolism in hypertrophic cardiomyocyte.Results:1. At 24 h and 48 h, compared with control, cells treated with AngⅡhad increased total protein content and enlarged diameter, but there were no difference between the two groups. At 72 h and 96 h, compared with control, total protein content and enlarged diameter were obviously increased.2. At 24 h, compared with control, myocardial cells treated with Ang II had increased MAO activity,but there was no difference between them. At 48 h.72 h,96 h,myocardial cells MAO activity were obviously or significantly increased. At 24 h and 48 h, compared with control, myocardial cell treated with AngⅡCOX had decreased activity, but there was no difference between them. At 72 h and 96 h, compared with control, myocardial cell COX activity was obviously or significantly decreased. At 24 h,48 h.72 h and 96 h, compared with control, myocardial cells treated with AngⅡhad increased the damage percentage of mitochondrial outer membrane. At 24 h,48 h,72 h and 96 h, compared with control, myocardial cells treated with AngⅡhad significantly reduced mitochondrial△Ψ. At 24 h,48 h,72 h and 96 h, compared with control, myocardial cells treated with AngⅡhad declined expression of Bcl-2, but there was no difference between the two groups. At 24 h, compared with control, myocardial cells treated with Ang II had declined expression of ANT, but there was no difference between them. At 48 h,72 h and 96 h, compared with control, the expression of ANT were obviously declined.3. At 24 h.48 h,72 h and 96 h, compared with control, myocardial cells treated with Ang II had obviously or significantly decreased the content of ATP,ADP,total adenosine acid pool and the level of energy charge. At 24 h, compared with control, myocardial cells treated with Ang II had increased the content of AMP, but there was no difference between the two groups. At 48 h.72 h and 96 h, compared with control, the content of AMP was obviously or significantly increased.4. At 72 h and 96 h, compared with control, ligustrazine and valsartan had significantly reduced myocardial cell total protein content, At 96 h, compared with control, radix astragali injection and shenfu injection had obviously reduced myocardial cell total protein content. At 72 h and 96 h, compared with control, ligustrazine, radix astragali injection and valsartan had obviously or significantly reduced cell diameter. At 72 h, compared with control, shenfu injection had obviously reduced cell diameter.5. At 72 h and 96 h. compared with control, ligustrazine. radix astragali injection and valsartan had obviously or significantly reduced myocardial cell MAO activity. At 72 h. compared with control, shenfu injection had obviously reduced myocardial cell MAO activity. At the two time points, compared with control, ligustrazine. shenju injection and valsartan had significantly increased mitochondrial COX activity. At 72 h, compared with control, radix astragali injection had obviously increased mitochondrial COX activity. At 96 h, mitochondrial COX activity was significantly increased. At the two time points, ligustrazine and valsartan had significantly reduced the damage percentage of mitochondrial outer membrane, radix astragali injection and shenju injection also had obviously reduced the damage percentage of mitochondrial outer membrane. At 72 h and 96 h, ligustrazine and valsartan had obviously improved mitochondrial AT compared with control. At 72 h, compared with control, radix astragali injection and shenju injection also had obviously improved mitochondrial△Ψ. At 96h, mitochondrial△Ψwas significantly improved. At the two time points, compared with control, ligustrazine,radix astragali injection,shenju injection and valsartan had increased expression of ANT. Bcl-2, but there was no differences between the two groups.6. At 72 h and 96 h, compared with control, ligustrazine, radix astragali injection and valsartan had significantly increased content of ATP. ADP. At 72 h, shenfu injection had obviously increased contents of ATP,ADP compared with control. At 96 h, contents of ATP. ADP were significantly increased. At the two time points, valsartan had significantly decreased content of AMP, ligustrazine, radix astragali injection had obviously decreased content of AMP. At 72 h, compared with control, shenfu injection had obviously decreased content of AMP. At 96 h, the content of AMP were significantly decreased. At 72 h, compared with control, valsartan had obviously increased content of total adenosine acid pool, radix astragali injection had significantly increased content of total adenosine acid pool. At 96 h, compared with control, shenfu injection had obviously increased content of total adenosine acid pool, ligustrazine. radix astragali injection and valsartan had significantly increased content of total adenosine acid pool. At the two time points, compared with control, ligustrazine,radix astragali injection. shenfu injection and valsartan had significantly increased the level of energy charge.7. At 72 h and 96 h, compared with control, trimetazidine had reduced myocardial cell total protein content, cell diameter, myocardial cell MAO activity, the damage percentage of mitochondrial outer membrane, increased expression of ANT, Bcl-2, but there were no differences between the two groups. At 72 h, compared with control, trimetazidine had obviously increased mitochondrial COX activity, mitochondrial△Ψ, contents of ATP. ADP. the level of energy charge. At 96 h, trimetazidine had obviously increased content of total adenosine acid pool compared with control. Conclusions:1. During the process of myocardial hypertrophy, damages of mitochondrial structure and function occurred.2. Changes of myocardial cell energy metabolism occurred in hypertrophic cardiomyocyte. Energy metabolism obstacle is a sensitive indicator in the early stage of myocardial hypertrophy.3. Ligustrazine. radix astragali injection,shenju injection and valsartan can restrain myocardial cell hypertrophy caused by AngⅡ, by protecting mitochondrial structure and function.4. Ligustrazine. radix astragali injection,shenju injection and valsartan can improve energy metabolism of the myocardial cell by restraining myocardial cell hypertrophy and protecting mitochondria.5. Trimetazidine had some good effects on energy metabolism in hypertrophic cardiomyocyte. Trimetazidine can't protect mitochondria and reverse myocardial cell hypertrophy.
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