Based On Network Pharmacology And Transcriptomics To Explore The Main Material Basis And Mechanism Of Taohong Siwu Decoction In Treating Ischemic Stroke

Objective:To further clarify the material basis of the Taohong Siwu Decoction(THSWD)by qualitative analysis of the in vivo and in vitro components.Through network pharmacology and transcriptomics sequencing technology to explore the treatment mechanism of ischemic stroke by THSWD.Method:UPLC-Q-TOF-MS~E combined with UNIFI data analysis platform was used to qualitatively analyze the main chemical constituents in the aqueous extract of THSWD.The rat model of middle cerebral artery occlusion(MCAO)was established and the THSWD were administered by gavage.Using UPLC-Q-TOF-MS~E combined with the in vitro chemical compositions of THSWD to identify the prototype components and metabolites after oral administration of THSWD.Based on the prototype components in vivo,predicting the targets of THSWD by network pharmacology and screening the targets related to ischemic stroke for GO analysis,KEGG pathway annotation.Then constructing a visualization network with components-targets-pathways.Through applying transcriptomics sequencing method to screen out the differentially expressed genes(DEGs)of THSWD in the treatment of ischemic stroke,and then combined with the results of network pharmacology prediction,the key targets and pathways for the mechanism of THSWD were selected for verification by animal experiments.Results:A total of 90 chemical components were identified in the aqueous extract of THSWD,including flavonoids,organic acids,glycosides,benzoquinones,etc.A total of 39 prototype components and 61 metabolites components were identified in MCAO rats,18 prototype components can enter the cerebrospinal fluid.A total of 437targets were predicted by network pharmacology with the in vivo components of THSWD,and 189 targets were related to ischemic stroke.There were 105 pathways by KEGG noted.High-throughput RNA sequencing technology was used to analyze the mRNA expression profiles of sham operation,MCAO and THSWD model in the left hemisphere of rats.A total of 1007 genes were identified.There were 16 enriched pathways(Homo sapiens)in the target genes.11 genes(Plau,Fabp4,Mmp9,Mmp12,Cfd,Lcn2,Trem1,Lgals3,Hmox1,Selp,Slc6a4)and 7 crossover pathways(Focal adhesion,Complement and coagulation cascades,Staphylococcus aureus infection,Malaria,Transcriptional misregulation in cancer,Progesterone-mediated oocyte maturation,PI3K-Akt signaling pathway)in the intersection of network pharmacology and transcriptomics.Animal experiments have proved that THSWD can reverse the abnormal expression of 11 target genes and inhibits the activation of the complement and coagulation cascades one of the 7 crossover pathways,therefore,THSWD can have the protection to against ischemic stroke.Conclusion:The UPLC-Q-TOF-MS~E analysis technology is combined with the UNIFI database analysis method which can quickly and comprehensively identify the in vitro and in vivo chemical components of THSWD.The main material basis for the treatment of ischemic stroke by THSWD was Mudanpioside E,paeoniflorin,Lactiflorin,Benoyylpaeoniflorin,?-Benzoyloxypaeoniflorin,etc.The mechanism may be related to the regulation of Complement and coagulation cascades,Cell cycle,Neuroactive ligand–receptor Interaction and other signaling pathways.The main material basis and mechanism of THSWD in treating ischemic stroke can be preliminarily clarified by using a combination of network pharmacology and transcriptomics.