| Objective: The in-hospital preparation Shuxin Shengmai Dan is an empirical formula based on more than 30 years of clinical experience.It has a definite effect on the treatment of cardiovascular diseases represented by coronary heart disease.In order to further explore its mechanism of action,this subject explores its protective mechanism for cardiomyocytes through analysis methods such as pathology and molecular biology,and explores its correlation with apoptosis,using experimental data to evaluate the efficacy of Shuxin Shengmai Dan,Shuxin Shengmai Dan's new drug development provides data support and provides valuable theoretical basis for the application of traditional Chinese medicine in the treatment of cardiovascular diseases and disease treatment.Method:1.70 healthy male SPF SD rats were randomly divided into 7 groups: blank group,sham operation group,positive control group(compound Danshen dripping pills),model group,Shuxin Shengmai Dan low dose group,Shuxin Shengmai Dan medium dose group,Shuxin Shengmai Dan high-dose group.2.Pretreatment of traditional Chinese medicine: After 5 days of adaptive feeding,they are administered in groups.Dosage: according to the "equivalent dose ratio of animal and human surface area conversion table" conversion,the Chinese medicine(Shuxinshengmaidan)middle-dose group is a healthy daily dose,the Chinese medicine(Shuxinshengmaidan)high-dose group(Shuxinshengmaidan)The low-dose group was divided into 2-fold dose and 0.5-fold dose,respectively,and the control group compound Danshen dripping pills was administered according to the daily dosage of healthy people.7days pre-administration,once a day,the blank group,sham operation group,model group were given distilled water.After 7 days,the model was fasted,the blank group was not treated,and the sham operation group was only threaded without ligation.The other groups underwent ligation of the left anterior descending coronary artery for 30 minutes and reperfusion for 120 minutes to prepare a rat model of myocardial ischemia-reperfusion injury.After successful modeling,whole blood and heart tissues were taken from the abdominal aorta.3.Monitor and record the changes of electrocardiogram and hemodynamics during ischemia/reperfusion,and analyze the effect of Shuxin Shengmai Dan on cardiac function of MI/RI model rats.4.HE staining and transmission electron microscopy were used to observe the myocardial tissue and ultrastructure of MI/RI rats,and the effect of Shuxin Shengmai Dan on MI/RI model myocardial tissue and ultrastructure was analyzed.5.The qRT-PCR method,Western blot method and IHC method were used to detect the mRNA and protein of SIRT1,p53,ac-p53,Bax,Bcl-2,Capase-3 and Capase-9 related factors of SIRT1 signaling pathway The amount of expression affects.To analyze the regulation of related factors on SIRT1/p53 signaling pathway of MI/RI rat myocardial tissue by Shuxin Shengmai Dan.Result:1.1.Electrocardiogram: After successful modeling,the electrocardiogram of each group of rats has obvious ST segment changes,and ST leads are elevated in the limb leads,and there are T wave inversion changes;after 30 minutes of reperfusion,the electrocardiogram of rats in each group suggests that the ST segment is different The change of the fall indicates that the MI/RI rats have been successfully modeled.2.Hemodynamics: Compared with the model group,the hemodynamic indicators of the rats in the Shuxin Shengmai Dan treatment group and the positive control group were improved to varying degrees,and the Shuxin Shengmai Dan group improved more obviously.3.HE staining: The MI/RI model group had incomplete myocardial tissue structure,disordered morphological arrangement,obvious interstitial edema,and shrinkage of the nucleus.Compared with the model group,the treatment groups of the Chinese medicine Shuxin Shengmai Dan were relatively complete in structure,with regular morphological arrangement and clear layers,and the degree of interstitial edema was reduced compared with the model group,and the nuclear shrinkage was reduced.Among them,Shuxin Shengmai Dan high-dose group showed the most obvious improvement.4.Electron microscopic observation: the muscle fibers of the model group were not arranged neatly,the mitochondria were swollen,and the arrangement was irregular.Somemitochondrial crests were incomplete and vacuoles appeared.The mitochondria of the Chinese medicine Shuxinshengmaidan treatment group and the positive drug group showed slight swelling and deformation,most of the structures were complete,and the muscle fibers were arranged roughly neatly,which was less than the model group.5.qRT-PCR method to detect the gene expression of SIRT1,Bcl-2,Bax,and Caspase-9in MI/RI rat myocardial tissue: Compared with the model group,Shuxin Shengmai Dan can up-regulate the gene expression of SIRT1,Bcl-2 The difference was statistically significant(P<0.05),and the Bax and Caspase-9 gene expression levels were reduced.The difference was statistically significant(P<0.05).Western blot detection of SIRT1,Caspase-9,p53,ac-p53 protein expression in myocardial tissue of MI/RI rats: Compared with the model group,Shuxin Shengmai Dan can up-regulate SIRT1 in MI/RI model rat The difference in the protein expression level was statistically significant(P<0.05),and the protein expression level of Caspase-9,p53,ac-p53 was down-regulated(P<0.05).IHC test indicated that Shuxin Shengmai Dan could up-regulate SIRT1 protein expression and reduce p53,Caspase-3 and Caspase-9 protein expression.Conclusion:1.Shuxin Shengmai Dan can improve the cardiac function and myocardial ultrastructure of rats after MI/RI,and has a protective effect on myocardial cells.2.Shuxin Shengmai Dan can regulate the expression of mRNA and protein of SIRT1,p53,ac-p53,Bax,Bcl-2,Capase-3,Capase-9 in myocardial cells of MI/RI model rats and reduce the occurrence of apoptosis,Its protective effect is related to the mechanism of apoptosis. |
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