| Objective: to explore the potential effect material basis and possible molecular mechanism of Zhenwu decoction in the treatment of heart failure(HF)by means of network pharmacology and molecular docking technology,to observe the therapeutic effect of Zhenwu decoction on the rat model of coronary artery ligation of heart failure through animal experiments,and to verify the potential biological process and molecular mechanism of network pharmacology,so as to further enrich the scientific connotation of Zhenwu decoction in the treatment of HF.It provides a scientific basis for the study of the modernization of Zhenwu decoction.Methods: 1.The chemical constituents and action targets of Zhenwu decoction were searched and screened by TCMSP,Pubchem,swiss ADME and other databases.The disease targets were obtained by searching the databases of Gene Cards,OMIM,Pharm GKB and Drug Bank.The intersection target of Zhenwu decoction and HF was obtained by drawing Venn diagram,and the potential active components and targets of Zhenwu decoction interfering with HF were obtained by constructing PPI and "drug-ingredient-target-disease" network.KEGG and GO enrichment analysis were carried out by using DAVID database,and the potential active components with the top 5 Degree values were docked with the target.two。 The rat model of heart failure after myocardial infarction was established by ligation of the anterior descending branch of the left coronary artery.4 weeks after operation,the surviving rats were randomly divided into 6 groups with 7 rats in each group:(1)sham operation group and model group were given normal saline for 28 days,(2)positive control group:losartan potassium 5mg/kg for 28 days,(3)Zhenwu decoction low dose group: Zhenwu decoction crude drug dose 6g/kg for 28 days.(4)Zhenwu decoction middle dose group: Zhenwu decoction crude drug dose 12g/kg for 28 days,(5)Zhenwu decoction high dose group: Zhenwu decoction crude drug dose 18g/kg for 28 days.The ECG of rats before and after modeling was observed,and the cardiac function and myocardial pathomorphological changes of rats in each group were observed by cardiac color ultrasound,LVMI,TUNEL detection,HE staining and serum BNP.The expression of apoptosis protein bax,bcl-2 and apoptosis key enzyme caspase3 in myocardial tissue of rats with heart failure was observed by immunohistochemical method,and the expression of PI3 K,AKT1 and p-AKT1 protein in myocardial tissue was detected by;Western Blot under different intervention conditions.Results: 1.68 chemical components,638 targets,1522 CHF disease targets,148 intersecting targets and 11 main active components of Zhenwu decoction were excavated.The key targets involved STAT3,IL6,EGFR,TNF,AKT1 and so on.KEGG analysis showed 122 pathways,which were closely related to PI3K-AKT signal pathway,c GMP-PKG signal pathway,HIF-1 signal pathway and so on.534 biological processes,121 molecular functions and 53 cellular components were obtained by GO analysis.Molecular docking showed that 6-gingerenol,β-sitosterol,kaempferol,3-epi-beta-Sitosterol and umbellate C had good binding activity with STAT3,IL6,EGFR,TNF,AKT1 and other targets.2)the ST segment of ECG in rats with coronary artery ligation was elevated,which indicated that the model of myocardial infarction was successful,2)the levels of LVMI and BNP in heart failure rats were significantly increased,while the levels of LVEF and LVFS were significantly decreased in heart failure rats,and the levels of LVMI,BNP,LVEF and LVFS were improved in low,middle and high dose groups of Zhenwu decoction(p< 0.01).3)HE staining showed that the myocardial fibers in the model group were disordered and broken,accompanied by cardiomyocyte swelling,hypertrophy and inflammatory cell infiltration,and the myocardial tissues of the other groups were relieved in varying degrees.TUNEL staining showed that the cardiomyocyte apoptosis in the model group was serious,and the apoptosis was alleviated after the intervention of Zhenwu decoction.4)compared with the sham operation group,the expression of Bax and Caspase3 in the model group increased significantly,the expression of Bax and Caspase3 decreased in the low,middle and high Zhenwu decoction groups,especially in the middle dose group,and the expression of Bcl-2 and the ratio of Bcl-2/Bax in the model group decreased significantly,especially in the middle and high dose groups.These results suggest that Zhenwu decoction may improve the degree of myocardial apoptosis in rats with heart failure by regulating the ratio of Bcl-2/ Bax and the expression of Caspase3.5)compared with the sham operation group,the expression of PI3 K,pAKT1 and AKT1 in the model group increased,and Zhenwu decoction could inhibit the expression of PI3 K,p-AKT1 and AKT1 protein.Conclusion: 1.Zhenwu decoction may have anti-HF effect through 6-gingerenol,β-sitosterol,kaempferol,3-epi-beta-Sitosterol and other components acting on PI3K-AKT,c GMP-PKG,HIF-1 α and other signal pathways.Each dose of Zhenwu decoction can reduce ventricular hypertrophy,enhance myocardial contractility and effectively improve cardiac function by inhibiting cardiomyocyte apoptosis.The mechanism may be related to the inhibition of PI3K-AKT1 pathway and the regulation of Bcl-2/Bax ratio and Caspase3 expression. |
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