The Investigation Of Benefiting Qi,Warming Yang,activating Blood And Promoting Diuresis On Prevention And Treatment In Chronic Heart Failure Based On Transcriptome And Network Pharmacology

Objective:The reasearch is aim to investigate the effects of Xinyang tablet in preventing and treating chronic heart failure.We analyzed the differential genes and enrichment pathways between Xinyang Tablet group,TAC model group and Sham group with transcriptomics technology.We concluded the key active ingredients,important target genes and main molecular mechanisms of Xinyang Tablet in the prevention and treatment of chronic heart failure by integrating transcriptomics technology and network pharmacology methods.Methods:1.Forty male C57 BL/6J mice were randomly divided into Sham group,TAC model group,traditional Chinese medicine group(Xinyang Tablet group)and positive control group(Perindopril group).The traditional Chinese medicine group was treated with Xinyang tablet,positive control group with Perindopril,while the others with saline.After eight weeks,we evaluated the Severity of heart pathology by echocardiographic testing and HE staining、Masson staining、Sirius red staining and WGA staining.2.After extracted the heart total RNA in Sham group、TAC model group and Xinyang Tablet group,we analyzed differential genes between groups,performed GO functional annotation and KEGG pathway enrichment in three groups by RNA-seq technology.3.We used three approachs to obtain the potential ative components of Xinyang Tablet:from HPLC-MS detection,filter ingredients with OB degree and DL degree in TCMSP website,from document records.We searched in Swiss Target Prediction platform for the prediction targets of these components.We filtered the intersection targets between network pharmacology prediction and transcriptome sequencing detection through Venny2.1.0,PPI network was established by STRING,GO functional enrichment and KEGG pathway enrichment analysis were performed with DAVID database.The drug-ingredients-targets-pathway network was constructed by Cytoscape to screen out key targets,core components and important pathways,the expression of key targets were tested and verified by fluorescent quantitative PCR.Results:1.As compared with the Sham operation group,the left ventricular ejection fraction(LVEF)、 left ventricular fractional shortening(LVFS)in model group were significantly decreased(P<0.01),the heart/weight ratio was significantly higher(P<0.01),and the cardiac collagen volume fraction was significantly higher(P<0.05).As compared with the model group,the LVEF 、 LVFS in Xinyang tablet group were significantly increased(P<0.01),the heart/weight ratio was significantly lower(P<0.01),and the cardiac collagen volume fraction was significantly lower(P<0.05).2.Within the transcriptome sequencing results,there were 1275 differential genes between the Xinyang tablet group and the TAC model group,the biological process changes between two groups were mainly enriched in the inflammation,immune response,cytokine secretion and regulation,blood circulation regulation,cardiac contraction,ion channels.The KEGG pathway were enriched in NOD-like receptors Signaling pathways,interactions between cytokines and cytokine receptors pathways,c GMP-PKG pathways,fatty acid metabolism pathways,adrenaline signaling pathways in muscle cells,AGE-RAGE in diabetic complications,cell cycle,p53 pathways.3.The main active ingredients in Xinyang tablets were quercetin、kaempferol、isorhamnetin、hederagenin、Jaranol、8-Isopentenyl-kaempferol、beta-sitosterol、Hypolaetin、Dinatin and 6-hydroxy-11,12-dimethoxy-2,2-dimethyl-1,8-dioxo-2,3,4,8-tetrahydro-1H-isochromeno[3,4-h]isoquinolin-2-ium,the important core targets were CDK1,ALOX5,PLK1,CCNB1,PDE5 A,CCND1,TNF,CCNA2,the main mechanisms were cell cycle,cell senescence,Fox O signaling pathway,PI3K-Akt signaling pathway,AMPK signaling pathway,and P53 signaling pathway.Conclusion:1.Xinyang tablet can effectively prevent and treat chronic heart failure by improving heart function,reducing myocardial fibrosis and hypertrophy.The core active ingredients of Xinyang tablet are quercetin 、 kaempferol 、isorhamnetin、hederagenin、Jaranol、8-Isopentenyl-kaempferol、beta-sitosterol、Hypolaetin、Dinatin and 6-hydroxy-11,12-dimethoxy-2,2-dimethyl-1,8-dioxo-2,3,2-4,8-tetrahydro-1H-isochromeno[3,4-h]isoquinolin-2-ium,these ingredients targeting CDK1,ALOX5,PLK1,CCNB1,PDE5 A,CCND1,TNF,CCNA2,the main effective mechannisms in the treatment of heart failureare are cell cycle,cell senescence,Fox O signaling pathway,PI3K-Akt signaling pathway,AMPK signaling pathway,and P53 signaling pathway.