Study On Molecular Mechanism Of Yiqi Huoxue Decoction In Treating Myocardial Ischemia Based On Network Pharmacology

Objective:To conduct computing network pharmacological studies on Radix Astragali seu Hedysari,Radix Angelicae Sinensis, Radix Ginseng, Radix Notoginseng and Rhizoma Ligustici Chuanxiong of Yiqi Huoxue Formula, in order to explore the active constituents in the treatment of myocardial ischemia, clarify the effective constituents of this formula in treating myocardial ischemia,in order to provide a research support for clarifying the pharmacodynamic material basis.Based onanimal models of myocardial infarction,usingthe early experimental the most significant effect Yiqihuoxue group to intervene,network pharmacological studies were applied to study pharmacological properties of compounds of YiqiHuoxue medicine and the related targets on myocardial ischemia.The useof echocardiography testing techniques investigate Yiqihuoxue drugs effect on NHE-1、CaMKⅡ、 VEGFR-2 and HIF-la expression of related genes and proteins.To use bioinformatics analysis of the effect of Yiqihuoxue drug, further reveal themolecular mechanisms of Yiqihuoxue drug.Methods:1. The collection and processing of the compounds contained in the YiqiHuoxue medicine: the chemical composition database was obtained by TCMD2009 database retrieval.The collection of target proteins related to myocardial ischemia:TTD database collected associated with myocardial ischemia disease target protein, Download all protein targets from the PDB database structure.2. On this basis, according to the target and target related diseases, the potential active components in the treatment of myocardial ischemia were predicted, and the material basis of Chinese herbal medicine was analyzed.3. Build an animal model of acute myocardial infarction by left coronary artery ligationsurgery and randomly divided into MI group, Yiqihuoxue group, Peiduopuli group, perindopril group,together with the shamoperated group. Second postoperative dayadministrated the rats in each group,respectively after 7 days,28 days get specimens.The use of echocardiography instrument detection of myocardial infarction after administration of 7 days,28 days left ventricular index, left ventricular volume, left ventricular diameter, left ventricular ejection fraction, left ventricular shortening fraction and left ventricular infarct border zone morphology cardiac index; the myocardial tissue structure was observed in the infarct area and the infarct border region.Using echocardiogram test for 7th day,28th day each time point cardiac function.Irrigate Yiqihuoxue medicine and western medicine to rats.Constrast related in different groups and in different time points.PCR to detect the expression ofCaMKI、NHE-1、VEGFR-2 and HIF-1α, CaMKIIprotein. Western Blotting to detect the expression of VEGFR-2 and HIF-1α.Results:1. Network pharmacological analysis results:If the degree more than 8,a total of 20 compounds had high network degree and betweeness with 20 anti-myocardial ischemia targets. Network analysis showed that YiqiHuoxue medicine played. A significant role in treating myocardial ischemia based on the synergistic effect of 20 active ingredients.VEGFR-2、NHE-1、CaMKⅡ、PKC are related to the strong function of the small molecule in the prescription ofYiqihuoxue medicine.2. The measurement results of each group of rats left ventricular function:MI groups ratsleft ventricular diastolic dimension(LVIDd) and left ventricular systolic dimension (LVIDs), left ventricular mass(LVM), left ventricular volume index of diastolic end(EDV)andleft ventricular volume index of shrink end(ESV), Ejection fraction (EF) and Fractional shortening(FS) compared with the shamgroupincreased at the 7thday,28thday time point,the difference was significantly(P<0.01), after the administration’s invention, administration groupLVIDscompared with the MIgroupwas significantlylower(P<0.05); MI groupFS and EFcompared with the shamgroup decreased; LVM, ESV, EDV, LVIDd and LVIDs were increased;MI group rats infarction marginal zone of myocardial cells arranged in disorder, texture direction uncertain, partially visible focal myocardial axons interrupt, a large number of inflammatory cell infiltration, myocardial fiber coloring and granular degeneration,but after drug intervention, myocardial cells could be seen in the myocardial cells arranged in disorder is not obvious, cytoplasm texture appeared a few irregular transverse band and a few inflammatory cells infiltrationin 28 days.3. Molecular biological indicator results:Molecular markers showed that the MI group rats compared with sham-operated group ratsof NHE-1 expression in infarct border zone myocardium and plasma were significantly increased (P<0.01), CaMKII mRNA levels were higher, after administration intervention, administration group NHE-1 concentration was significantly lower than the model group, CaMKII mRNA expression levels decreased, with statistical significance (P<0.01).After 7 days modeling, treatment group compared withMI group, VEGFR-2 and HIF-la gene and protein expression were increased and had significant difference (P< 0.01); and in 28 days, MI group compared in sham operation group, the expression of myocardial tissue of rats in model group mRNA gene and protein VEGFR-2 significantly increased, there was a significant difference (P<0.01). Compared with the model group, the expression of mRNA VEGFR-2 gene and protein in the other three groups were decreased, and the difference was statistically significant (P<0.01).Analysis conclusion:1. Network pharmacology research methodsis helpful to analyze the possible active molecules in the prescription ofYiqihuoxue active molecules, and to retrievethe active ingredient treatment of myocardial ischemia disease related targets.2. Yiqihuoxue Formula can improve the blood supply of myocardial infarct border zone, in order to achieve the effect of improving myocardial ischemia and left ventricular function of cardiac structures, when hypoxia of ischemic, the body of sodium hydrogen compensatory exchange reaction, and by interve Yiqihuoxue Formula,it can regulate NHE-1, the gene expression of CaMKII, suppressing infarct border zone of myocardial tissue cells calcium overload, to improve myocardial tissue vasodilatio abilitys, increased myocardial blood flow, and thus it is possible to improve myocardial ischemic symptoms; at the same time through the study found Yiqihuoxue Formula can promote the expression of VEGFR-2 and HIF-la gene and protein, suggesting Yiqihuoxue Formula may be able to promote angiogenesis and maturation of myocardial ischemia, ischemic make myocardial tissue blood supply is improved, thereby slowing the course of myocardial ischemia in the direction of poor development.Innovation:1. In this study, we combined network technology and pharmacology animal experiments, to explain the Yiqihuoxue Formula treatment of myocardial ischemia mechanisms at the molecular level with the traditional Chinese medicine theory,as well as provide a scientific basis to Yiqihuoxue Formula in multi-components,multi-targets, multi-channels of pharmacodynamic mechanism.2. This study clarified that Yiqihuoxue Formula can promote myocardial tissue of myocardial ischemia rats in blood vessels newborn and mature.At the same time, it can be adjusted by calmodulin kinase signaling pathways to regulate gene expression of CaMKⅡ NHE-1, and it may protect ischemic myocardial cells, thereby improving cardiac dysfunction due to ischemic myocardium.