| Background and purpose After myocardial infarction,oxidative stress-induced cardiomyocyte apoptosis is a main cause of ventricular remodeling,and oxidative stress-induced mitochondrial autophagy is also involved in the pathogenesis of myocardial infarction,but it work as duality by myocardial protecting or injuring.According to the traditional Chinese medicine theory of the Qi and blood the Yiqihuoxue Decoctionwas created,which is the clinical treatment of myocardial ischemic experience formula.Previous studies have shown that Yiqi Huoxue Decoction can effectively improve the clinical symptoms and cardiac function of patients with myocardial infarction and improve coronary microcirculation.Therefore,this study aims to explain the pharmacological mechanism of Yiqi Huoxue Decoction on myocardial injury after cardiomyocyte apoptosis and mitochondrial autophag.MethodThe animals were randomly divided into 4 groups:MI group,Yiqi Huoxue group and Perindopril group,and the sham operation group.The rats in each group were treated with the corresponding drugs on the 2nd day after operation.The changes of the related indexes were observed at 7 and 28 days after treatment.The cardiac function and structural changes of myocardial infarction rats were measured to evaluate the effect of Yiqi Huoxue Decoction on cardiac function in rats with myocardial infarction.Microscopic is used to observe myocardial cell structure and transmission electron microscopy(TEM)is used to observe the mitochondrial structure,morphological changes and the number of autophagic vesicles in rats with myocardial infarction,which is to investigate the protective effect of Yiqi Huoxue Recipe on myocardium of myocardial infarction rats and its effect on myocardial mitochondrial autophagy.To investigate the changes of serum superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)in rats with myocardial infarction by double antibody sandwich enzyme-linked immunosorbent assay(ELISA),to investigate the impact of Yiqi Huoxue Recipe onoxidation capacity.The levels of LC3 and Beclinl mRNA and protein were detected by Western Blotting and real-time fluorescence quantitative(RT-PCR)in the marginal zone of myocardial infarction.To investigate the mechanisms of Myocardial Mitochondrial Autophagic Changes and Qi and Activating Blood Circulation in Rats.The expression of Cyt c and Caspase-3 protein were observed by Western Blotting.Evaluate the changes of myocardial cell apoptosis and the effect of Qi The effect of blood circulation on it.ResultsThe experimental study consisted of five partsReplication of rat model of myocardial infarction,conpare heart structure and function changes in at 7 days and 28 days.Compared with Sham group,LVEF and LVFS were declined,and LVIDs,LVIDd,EDV,ESV were increased in myocardial infarction group.At 7 days,compared with myocardial infarction group,rat gived Yiqi Huoxue Recipe and Perindopril were increased in LVEF,LVFS apparently,LVIDs,LVIDd,EDV,ESV were declined but no statistically significant.At 28 days,compared with myocardial infarction group,rat gived Yiqi Huoxue Recipe and Perindopril were increased in LVEF,LVFS,declined inEDV,ESV apparently.It shows that left ventricular dilatation,systolic and diastolic function decreased after myocardial infarction,Yiqi Huoxue Recipe can release left ventricular dilatation,Improve left ventricular systolic and diastolic function.Effects of Yiqi Huoxue Recipe on Myocardial Tissue and Mitochondrial Morphology in Myocardial Ischemic Model Rats:The light microscopy showed that the myocardial cells in the sham operation group were neatly arranged at 7 days,the texture was clear,muscle fibers were clear and no obvious inflammatory cell infiltration was found.The myocardial cells in the myocardial infarction group were disordered and the texture was uncertain.The myofibroblasts were disrupted and the nuclei were dissolved and disappeared.Compared with the model group,the pathological changes of the perindopril group were similar to those of the model group,but the range of the lesion was smaller than that of the model group.At 28 days,the myocardium of the sham operation group was neatly arranged,the texture was clear,the structure of the myocardial fiber was clear,and no myocardial fibers and collagen tissue were observed.The myocardial cells in the myocardial infarction group were disordered and the texture was uncertain.In the perindopril group,the proliferation of cardiomyocytes was obvious,and the fibroblasts were proliferated in some parts,and the range of lesion was decreased.Electron microscopy showed that the mitochondria were vacuolated at 7 days,and the typical autophagosomes and autophagosomes were formed.A small amount of autophagy was observed in the drug group,which was lower than that in the model group.At 28 days,the number of mitochondria in the model group was abnormally hyperplastic,morphological disorder,vacuolization was obvious and the autophagic bubble was rare.The mitochondrial structure of the drug group was clear and the phagocytosis was obvious.The results showed that the myocardial cell death,interstitial fibrosis and other pathological changes after myocardial infarction,myocardial mitochondria morphological structure damage,autophagy increased with time gradually decreased.Yiqihuoxue decoction can reduce myocardial cell death and interstitial fibrosis,reduce myocardial cell mitochondrial structure damage,and increase with time to increase the phenomenon of autophagy.To investigate the effects of Yiqi Huoxue Recipe on SOD and GSH-Px in serum of rats with myocardial ischemia model:SOD and GSH-P in the model group were significantly lower than those in the sham operation group,The difference was significant.Compared with model group,SOD and GSH-PX in Yiqi Huoxue group and Perindopril group were significantly higher than those in model group.The results showed that after myocardial infarction,rat antioxidant capacity decreased,the use of Yiqihuoxue can increase the rat antioxidant capacity.To investigate the effect of Yiqi Huoxue Recipe on the expression of autophagy-related molecules LC3,Beclinl gene and protein in myocardium of myocardial infarction rats in infarcted area.The expression of LC3,Beclinl gene and protein in myocardial tissue of rats with myocardial infarction was significantly higher than that in sham operation group.The expression of LC3 and Beclinl were significantly increased in myocardial infarction group.Compared with the rats with myocardial infarction,the expression of LC3 gene in Yiqi Huoxue group decreased,but there was no significant difference.The expression of Beclinl gene was decreased and the expression of LC3 and Beclinl gene in perindopril group decreased.At 28 days,the expression of LC3 and Beclinl was significantly increased in myocardial infarction group compared with sham operation group.Compared with the rats with myocardial infarction,the expression of LC3 and Beclinl gene in Yiqi Huoxue group and perindopril group were significantly increased.The expression of LC3A,LC3B and Beclinl protein in myocardial infarction group were significantly higher than those in sham operation group at 7 days.Compared with the rats with myocardial infarction,the expression of LC3A,LC3B and Beclinl protein in Yiqi Huoxue group and perindopril group were significantly decreased.At 28 days,the expression of LC3A and LC3B protein in myocardial infarction group were significantly higher than those in sham operation group.Compared with the rats with myocardial infarction,the expression of LC3A,LC3B and Beclinl protein in Yiqi Huoxue group and perindopril group were significantly increased.The results showed that the expression of autophagy protein and gene increased at 7 days after myocardial infarction,and decreased at 28 days.The use of Yiqi Huoxue Decoction could regulate the autophagy protein and gene expression in cardiomyocytes at 7 and 28 days,that is,Increase day by day.To investigate the effect of Yiqi Huoxue Decoction on the expression of Cyt c and caspase-3 protein in myocardium of myocardial infarction model in rats with myocardial ischemia.The expression of CytC and caspase-3 protein in model group was significantly lower than that in model group Group was significantly increased,drug expression decreased after drug intervention.The expression of CytC and caspase-3 protein in the model group was higher than that in the sham operation group at 28 days,but there was no significant difference after drug intervention.The results showed that after myocardial infarction,the apoptotic phenomenon was more significant at 7 days,and the use of Yiqi Huoxue Decoction could relieve myocardial cell apoptosis.28 days although the phenomenon of apoptosis is not significant,but Yiqi Huoxue side still show the role of reducing the role of myocardial cell apoptosis.Conclusion1 Yiqi Huoxue Decoction can effectively improve the cardiac structure and function changes in rats with myocardial infarction,improve the antioxidant capacity of myocardial cells in rats,and inhibit the early cardiomyocyte apoptosis after myocardial infarction.2 Cardiomyocyte mitochondrial autophagy changes with time node,Yiqi Huoxue Decoction at different time nodes were inhibited and promote autophagy,worked in myocardial protection. |
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