Study On The Mechanism Of Zhishi Xiebai Guizhi Decoction In The Treatment Of Coronary Heart Disease Based On Network Pharmacology

Objective: Using network pharmacology and bioinformatics analysis methods to study the key drug components,targets and potential mechanism of action of Zhishi Xiebai Guizhi Decoction in the treatment of coronary heart disease.In vivo and in vitro experiments were conducted to verify the curative effect of the prescription and the molecular biological mechanism of its key components in treating coronary heart disease.Methods:1.Using Drug Bank,Genecards,OMIM,TTD,and Dis Ge NET databases to obtain coronary heart disease-related genes,and using TCMSP and Swiss Target Prediction databases to obtain the main active ingredients of Zhishi Xiebai Guizhi Decoction and predict their targets.Use R language to conduct GO,KEGG and DO enrichment analysis on the target genes in the intersection part,and perform topology analysis on the Chinese medicine component-target network and protein interaction network to find key drug components and key targets.2.A mouse model of coronary heart disease was constructed by injecting PCSK9 adeno-associated virus into the tail vein and fed with high-fat diet,and treated with Zhishi Xiebai Guizhi Decoction by gavage for 1 month,and performed aortic oil red O staining,blood lipid detection,and RT-PCR quantification,to evaluate the curative effect of Zhishi Xiebai Guizhi Decoction,and to verify the screened key targets.3.Using oxidized low-density lipoprotein to intervene human umbilical vein endothelial cells to establish a model of atherosclerotic functional injury.Western blotting was used to observe the expression of apoptosis-related proteins(Bcl-2,Bax)and PI3K-Akt-Bax signaling pathway-related proteins after Naringenin intervention.Results:1.Through network pharmacological analysis,6253 genes of coronary heart disease,51 active components of traditional Chinese medicine and 827 drug targets were obtained,and 50 key targets were screened out after topological analysis.GO was enriched to 3691,of which 3209 were enriched in BP,137 in CC and 345 in MF.BP is mainly enriched in biological processes such as positive regulation of MAPK cascade,response to oxidative stress,peptide tyrosine phosphorylation and modification,calcium homeostasis,vascular process of circulatory system and so on.KEGG is mainly concentrated in PI3K-Akt,lipid and atherosclerosis,FOXO,insulin resistance and endocrine resistance and other signal pathways.Through molecular docking,it was proved that naringin could stably bind to the protein receptors of PI3 K,Akt,Bcl-2and Bax.2.Zhishi Xiebai Guizhi Decoction can significantly reduce vascular atherosclerotic plaque(P<0.05),and this effect is concentrationdependent.It can significantly reduce the level of triglyceride and increase the level of high-density lipoprotein cholesterol(P<0.05).Although the level of LDL-C is lower than before,there is no statistical significance(P>0.05).Zhishi Xiebai Guizhi decoction can up-regulate the expressions of PIK3 CA,Akt1,SRC and STAT3(P<0.05).3.The appropriate concentration of naringenin to intervene human umbilical vein endothelial cells is 40 μmol/l.Naringenin can up-regulate The expression of PI3 K,p-PI3 K,p-Akt and Bcl-2 inhibited the expression of Bax(P<0.05),and increased Bcl-2/Bax(P<0.05).Conclusions:1.Zhishi Xiebai Guizhi Decoction can reduce triglyceride and Low density Lipoprotein cholesterol,increase High density lipoprotein cholesterol,and reduce atherosclerotic plaque formation.2.Naringenin,β-sitosterol,and quercetin may be the key components of Zhishi Xiebai Guizhi Decoction,which can regulate the expression of PIK3 CA,Akt1,SRC,and STAT3 to play a role.3.Naringenin may inhibit ox LDL-induced apoptosis of HUVECs by activating the PI3K-Akt-Bax signaling pathway.