| Purpose: Using network pharmacology methods,to explore the predictive targets and mechanism of action of Astragalus compound Chinese medicine in the treatment of coronary heart disease by using the intersection target gene formed by the target gene of Astragalus compound Chinese medicine and the target gene of coronary heart disease.Through cell culture technology,the subjects selected rat myocardial microvascular endothelial cells(RCMECs)to observe the interventional effect of Astragalus compound Chinese medicine serum on homocysteine-induced RCMECs injury and the mechanism of inflammation,and further explore the treatment of Astragalus compound Chinese medicine The mechanism of coronary microcirculation disorder,broadening the treatment of coronary microcirculation disorder,also provides theoretical experimental basis for the clinical application of Astragalus compound Chinese medicine in traditional Chinese medicine.Materials and methods:(1)Based on network pharmacology,search the Astragalus compound Chinese medicine components and target genes and coronary heart disease target genes through the database,take the intersection of the two target genes,and use the PPI network and DAVID 6.8 database to analyze the intersection target genes Reflect the key target genes and the biological processes involved in the intersection target genes,etc.,explore the possible mechanism of Astragalus compound Chinese medicine in the treatment of coronary heart disease;use Cytoscape 3.6.0 software to analyze "Astragalus compound-Chinese medicine-components-intersection target genes" "Network" conducts network topology analysis to clarify the key components of Astragalus compound Chinese medicine.(2)According to the needs of the experiment,50 male SPF SD rats were randomly divided into five groups according to the standard of 10 rats.They were named blank group,model group,low-dose astragalus compound Chinese medicine group,and astragalus Compound Chinese medicine middle-dose group,Huangqi compound Chinese medicine high-dose group,blank group and model group: the same volume of distilled water was given to the stomach,the astragalus compound Chinese medicine low-dose group was administered according to 1.8 g/kg,and the astragalus compound Chinese medicine middle-dose group was according to 3.6 g/kg standard,high-dose astragalus compound Chinese medicine group was according to 7.2 g/kg standard.Rats in five groups were given intragastric administration twice a day for 7 consecutive days.On the 7th day,the five groups of rats after 1 hour of intragastric treatment were anesthetized first,and then blood was collected through the abdominal aorta.The collected serum was first allowed to stand,then centrifuged at a high speed,and finally filtered.Inactivate,and finally put the processed drug-containing serum in a refrigerator at-80 ? for subsequent use.(3)The purchased rat myocardial microvascular endothelial cells were subcultured,frozen and resuscitated,and the morphology was observed under an inverted microscope.The positive rate of the cells identified by CD31immunofluorescence method was greater than 95% for use.The cells were divided into blank group,model group,low,medium,and high dose groups of Astragalus compound Chinese medicine.All groups except the blank group were given homocysteine to induce cell damage.Astragalus compound Chinese medicine low,medium and high dose groups were respectively Give the corresponding Astragalus compound medicated serum low,medium and high dose medicated serum to co-culture with the cells for 24 hours,and collect the cell supernatant of each group to facilitate the determination of indicators.For cell viability detection,the MTT method can be used Detection,for the detection indicators TNF-? and IL-6,the content of both can be detected by the ELISA method.Result:1.After the data retrieval,screening,sorting and summarizing in the early stage of this research,it is concluded that the four Chinese herbal medicines of Astragalus,Astragalus,Angelica,Scorpion,and Centipede contain 39 active ingredients of Chinese medicine,which correspond to 326 target genes.Heart disease contains 1024 target genes.Using network pharmacology to study the relationship between Astragalus compound Chinese medicine and coronary heart disease,it can be seen that the 30 active ingredients of Astragalus compound Chinese medicine correspond to 94 intersection target genes.The results of PPI network topology analysis and enrichment analysis of the intersection target genes show that the inflammatory response is the key to the entire biological process,and there are key target genes such as IL-6 and TNF involved in the inflammatory response,which may be astragalus The mechanism of compound Chinese medicine in treating coronary heart disease.2.Analyze and compare the contents of IL-6 and TNF-? in the supernatant of the five groups of RCMECs by ELISA method: Compared with the blank control group,the contents of IL-6and TNF-? in the cell supernatant of the model group were significantly increased(P<0.01),compared with the model group,the contents of IL-6 and TNF-? in the supernatant of RCMECs in the high-dose astragalus compound Chinese medicine group,the middle-dose astragalus compound Chinese medicine group,and the low-dose astragalus compound Chinese medicine group were significantly reduced(P<0.01).Conclusion:1.Through network pharmacological analysis,Astragalus compound Chinese medicine can treat coronary heart disease through key targets such as IL-6 and TNF and multiple pathways.2.Astragalus compound Chinese medicine-containing serum can significantly reduce the levels of IL-6 and TNF-? inflammatory factors in the cells of the experimental group,and reduce the damage of Hcy to rat myocardial microvascular endothelial cells,which may be one of the mechanisms of its treatment of coronary microcirculation disorders First,there is a dose-effect relationship between the astragalus compound Chinese medicine groups. |
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