| 1 BackgroundChronic heart failure is the end and severe stage of various heart diseases,which is a common chronic cardiovascular disease in the world today.Some data show that the 5-year mortality rate of hospitalized patients with heart failure reaches 50%,which has greatly affected human society.Ginseng is a common kind of Traditional Chinese Medicine which can be used to regulate blood pressure,recover heart function and relieve physical weakness.The research team used integrative pharmacology and molecular docking techniques in the early stage and found that the efficacy of ginseng supplementing qi may be related to intervention in human energy metabolism-related pathways,while chronic heart failure is also closely related to human energy metabolism process.Based on the above considerations,this study starts with the pharmacodynamic evaluation and molecular mechanism of ginseng for chronic heart failure,and provides a certain theoretical basis for the clinical application of ginseng and the molecular mechanism of chronic heart failure.2 Methods and results2.1 Pharmacodynamic evaluation of ginseng on chronic heart failurePharmacodynamic evaluation was carried out in this section by the chronic heart failure model induced by myocardial infarction caused by the left anterior descending coronary artery ligation.According to the characteristics of this model,the time to develop to chronic heart failure after myocardial infarction caused by ligation of the left anterior descending coronary artery is about 4 weeks.Therefore,rats are fed normally for 4 weeks after modeling.After that,Doppler ultrasound was used to test the cardiac function of rats,and the successfully modeled rats were selected for follow-up experiments based on rat left ventricular ejection fraction(EF%).Firstly,rats were divided into 5 groups randomly including ginseng group,positive drug(valsartan group),sham operation group,model group and control group.Doppler ultrasound test is performed every 2 weeks,mainly examined the left ventricular ejection fraction(EF%),left ventricular short-axis shortening rate(FS%),left ventricular posterior wall end-diastolic thickness(LVPWd),and left ventricular posterior wall end-systole thickness(LVPWs),left ventricular end-diastolic diameter(LVIDd),left ventricular end-diastolic diameter(LVIDs)to initially evaluate the efficacy of the ginseng drug.Then the Pressure-Volume(PV)-Loop system was used to further detect the heart function of rats.Subsequently,enzyme-linked immunosorbent assay(ELISA)was used to detect brain natriuretic peptide(BNP),a gold measure of heart failure,and combined the effects of HE on pathological sections to evaluate the efficacy.The rats were sampled after the administration for 6 weeks.Four weeks after the rats were modeled,the.EF%of rats ware significantly decreased tested by ultrasound.The animals were randomly divided into 5groups including:model group,ginseng group,valsartan group,sham operation group and control group.Doppler ultrasound test is performed every 2 weeks and samples were taken at 6 weeks.Left ventricular ejection fraction(EF%),left ventricular short axis shortening rate(FS%),left ventricular posterior wall end-diastolic thickness(LVPWd),left ventricular posterior wall end-systolic thickness(LVPWs)were significantly higher in concentration group than those in the model group;left ventricular end-diastolic diameter(LVIDd)and left ventricular end-diastolic diameter(LVIDs)of the ginseng group and the valsartan group were significantly lower than those in the model group.The PV-Loop testing showed that the EF%of rats was significantly improved after administration.Brain natriuretic peptide(BNP),a golden indicator of heart failure,was significantly lower in ginseng group rats and valsartan group than in model group.Myocardial tissue and cardiac function were improved in ginseng group and valsartan group rats showed by the HE pathological section.These provide the evidence that ginseng and valsartan can protect the heart and relieve heart failure to a certain extent.2.2 Study on the molecular mechanism of ginseng acting on chronic heart failureIn this section,based on the results of previous experiments and research in the literature,the representative left ventricular sample from model,ginseng and sham group was used to perform second RNA-Seq generation sequencing to screen out the differentially expressed genes between 3 groups,then the functional enrichment analysis of differentially expressed genes and detection of the related differentially expressed genes were performed to explore possible mechanisms of ginseng acting on chronic heart failure.The differentially expressed genes were selected according to the screening criteria which is satisfying false discovery rate FDR<0.05 and |Fold change|>1.5.Then the KEGG pathway enrichment analysis was performed on the differential genes.Some genes involved in oxidative phosphorylation such as Ndufa4,Atp5pd,Atp5mc3,Atp5fla,Sdhb and Cox10,some genes involved in fatty acid metabolism such as Echsl,Acsl6,Acsll,Acoxland some other genes involved in PPAR signaling pathway such as Acadl,Acsl6,Acsll,Acox1 showed a certain degree of callback after administration.This indicates that ginseng may play an intervention role in heart failure by intervening in related energy metabolism pathways.After that,the related pathways were enriched by the 400 up-regulated genes after ginseng administration,30 KEGG pathways including tricarboxylic acid cycle,oxidative phosphorylation,propionate metabolism,pyruvate metabolism,fatty acid biosynthesis,carbon metabolism,fatty acid degradation,fatty acid metabolism,lysine degradation and PPAR signaling pathways were enriched.While the 23 related pathways enriched by 327down-regulated genes including cytokine-cytokine receptor,Ras signaling pathway,NF-?B signaling pathway,Staphylococcus aureus infection,chemokine signaling pathway.The results of this study show that ginseng can improve heart failure by enhancing related energy metabolism processes and reducing the expression of inflammation-related factors to a certain extent.The detection the differentially expressed gene PPAR ? and its downstream factors Acox and Acsll further indicated that ginseng can enhance energy metabolism processes in the body.The results of this study indicate that ginseng can play a role in heart failure by enhancing processes related to energy metabolism in the body to a certain extent.2.3 Study in vitro of ginseng's intervention on chronic heart failureBased on the results of the RNA-Seq and literature survey,the key pathways and the relationships between these pathways was analysed,then the key transcription factors was selected for vitro experiments.A variety of ginsenoside components of ginseng have been shown to play a role in treating chronic heart failure,in order to further confirm whether ginsenosides are the major components in the treatment of chronic heart failure in ginseng,the total ginsenosides in the ginseng extraction was extracted.The H9c2 cells were selected as the research objects,and the ginseng extraction,total ginsenosides were given to normal cultured cells respectively.The changes of key transcription factor were observed.The effect of ginseng on chronic heart failure was further studied in vitro based on animal experiments.Based on the results of the RNA-Seq and literature survey,ginseng may exert a therapeutic effect on heart failure by affecting the energy metabolism process.Among the enriched energy metabolism pathways,the lipid metabolism pathway is upstream of some energy metabolism pathways,and PPAR ? is a key factor of the PPAR signaling pathway in lipid metabolism.Therefore,we chose PPAR ? as the key transcription factor for research and the ginseng extraction and total ginsenosides were given to normal cultured cells respectively to test the gene expression of PPAR?.The results showed that the expression of PPAR ? gene increased to some extent.3 ConclusionThis study proves that ginseng can release the disease and development process of chronic heart failure induced by myocardial infarction caused by ligation of left anterior descending coronary artery to a certain extent,and it has a certain degree of improving cardiac function.Second RNA-Seq generation sequencing technology was used to analyze the(DEGs)between different 3 groups,it was found that after administration of ginseng,it may play a certain role in regulating the PPAR signal pathway,tricarboxylic acid cycle,and fatty acid metabolism.The possible mechanism for the treatment of heart failure is to intervene in the body's energy metabolism-related processes.Based on the results of the RNA-Seq and literature survey,the key transcription factor PPAR ? were selected for cell research.It was found that PPAR ? has been up-regulated to a certain extent after ginseng extraction and total ginsenosides were given to normal cultured cells respectively,which further proves that ginseng's intervention in heart failure may be related to the process of energy metabolism,and it's therapeutic effect on heart failure may be achieved through the intervention on PPAR signaling pathway. |
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