| Objective:Qi deficiency liver cancer(QDLC)is a common malignancy and an important part of liver cancer research in traditional Chinese medicine(TCM).In the course of its treatment,Panax ginseng is often selected as the main Chinese herbal medicine,and its function has special significance in the tumor treatment of Qi deficiency constitution.At present,there are many researches on Panax ginseng in the treatment of tumors in modern medicine,but there are few studies on QDLC,and the related mechanism is not clear.Therefore,this study is mainly based on the pharmacodynamic evaluation of the effective parts of Panax ginseng in the treatment of QDLC,the effects of effective parts of Panax ginseng on endogenous substances in the animal model of QDLC,investigation of the metabolic regularity of ginsenosides in the animal model of QDLC and the study of the interaction between different effective parts of Panax ginseng,to elucidate the therapeutic effect of Panax ginseng on QDLC and the related mechanism,and to provide experimental basis and data support for the clinical application of Panax ginseng in TCM.Methods:1.The animal model of QDLC was optimized and established by swimming exhaustion and xenograft Hep G2.The model of qi deficiency was evaluated and explained from the point of view of biomarkers and metabolic pathway by using UPLC-Q-TOF/MS metabolomics method,combined with the appearance of physical signs.The effect of qi deficiency on the occurrence and development of liver cancer was investigated by observing the formation and development of qi deficiency and tumor,blood routine,histopathology and serum metabonomics.2.According to the methods of"Panax ginseng"and"total ginsenosides"in the Pharmacopoeia of the people’s Republic of China,combined with literature,the preparation method of effective parts of Panax ginseng were optimized,and ginsenosides were identified and analyzed by using ultra-high performance liquid chromatography and tandem quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF/MS).Then the model was intervened with ginseng extract,ginsenosides,polysaccharides and the mixture of ginsenosides and polysaccharides(1:1).And through the observation of tumor development process,tumor inhibition rate,organ index,blood routine,histopathology and immunohistochemistry,the therapeutic effect and synergistic effect of effective parts of Panax ginseng on QDLC were investigated.3.UPLC-Q-TOF/MS metabonomic analysis,principal component analysis(PCA),orthogonal partial least squares discriminant analysis(OPLS-DA)and MS/MS mass spectrometry tandem technique were used to investigate the effects of effective parts of Panax ginseng on endogenous metabolites in urine and serum of QDLC model.At the same time,the pharmacodynamic potential biomarkers of the effective parts of Panax ginseng and the synergistic potential biomarkers of ginsenosides and polysaccharides on QDLC were screened and identified,and the metabolic pathways related to the biomarkers were investigated.4.Fecal metabonomics and 16S r RNA sequencing techniques were used to investigate the changes of fecal endogenous metabolites and gut microbiota during the treatment of QDLC with the effective parts of Panax ginseng.Then,the relationship among host phenotype,gut microbiota and fecal metabolites was comprehensively analyzed by spearman correlation coefficient.5.Through the investigation of the components of Panax ginseng absorbed into the blood of the QDLC model,the exploration of the correlation between pharmacokinetics and pharmacodynamics(PK-PD)and the analysis of targeted network pharmacology,the interaction and response between the model and Panax ginseng were expounded,and the material basis and whole action mechanism of Panax ginseng in the treatment of QDLC were investigated.Results:1.Through the investigation of the establishment of QDLC model,the best method for establishing the model was determined as follows:fixing 5%body weight metal in the tail to swim until exhaustion(nose tip immersed in water>5s)for 15 days to establish Qi deficiency model.After the establishment of Qi deficiency model,Hep G2 cell suspension was inoculated subcutaneously into the axilla(the cell concentration was 5.0×10~7/ml,and each mice was inoculated with 50μL).Through the investigation of urine metabonomics,combined with the physical signs and growth status of tumors,the QDLC model was successfully evaluated,and it was also verified that there was a close correlation between qi deficiency and energy metabolism.In addition,through the study of the effect of qi deficiency on the occurrence and development of liver cancer,it was found that qi deficiency could promote the development of tumor,affect the physiological and pathological state of the body,and caused endogenous substance disorder.2.Through the optimization of preparation methods,the best preparation methods of effective parts of Panax ginseng were determined,which the main technical routes were water extraction,alcohol precipitation and D101 macroporous resin separation.Then,40kinds of ginsenosides were identified by UPLC-Q-TOF/MS.The results of pharmacodynamic evaluation showed that the effective parts of Panax ginseng improved the appearance of signs,inhibited the growth of tumor,regulated the changes of blood routine and histopathology and the expression of Bax,Bcl-2 and VEGF-B in the treatment of QDLC,indicating that all effective parts of Panax ginseng had good therapeutic effect on QDLC,and ginsenosides and polysaccharides might have synergistic effect.3.Through the study of urinary metabonomics,38 pharmacodynamic potential biomarkers of the effective parts of Panax ginseng and 6 synergistic potential biomarkers of ginsenosides and polysaccharides on QDLC were screened and identified,and 11 metabolic pathways related to the biomarkers were investigated,including purine metabolism,phenylalanine metabolism,arachidonic acid metabolism,citrate cycle and so on.Through the study of serum metabonomics,13 pharmacodynamic potential biomarkers of the effective parts of Panax ginseng and 7 synergistic potential biomarkers of ginsenosides and polysaccharides on QDLC were screened and identified,and 7 metabolic pathways related to the biomarkers were investigated,which mainly included purine metabolism,phenylalanine metabolism,arachidonic acid metabolism,glycerolipid metabolism,unsaturated fatty acid biosynthesis,sphingolipid metabolism and steroid hormone biosynthesis.4.Through the investigation of endogenous substances in feces,31 pharmacodynamic potential biomarkers and 20 synergistic potential biomarkers of effective parts of Panax ginseng on QDLC were screened and identified.And then,metabolic pathways related to the biomarkers were investigated,which mainly included bile acid biosynthesis,unsaturated fatty acid biosynthesis,tryptophan metabolism,arachidonic acid metabolism,pyrimidine metabolism,vitamin B6 metabolism and so on.In the study of gut microbiota,at the genus level,25 species of bacteria with significant differences of effective parts on QDLC and 23species of bacteria with significant differences of synergistic action of ginsenosides and polysaccharides were screened.In addition,Spearman correlation analysis showed that there was a complex potential relationship among phenotype,gut microbiota and fecal metabolites during the development of QDLC and Panax ginseng intervention.5.The results of material basis analysis showed that a total of 15 ginsenosides were absorbed into the blood of model animals when Panax ginseng was used to treat QDLC.The results of PK-PD correlation study showed that the endogenous substances changed clockwise or counterclockwise with the concentration of active components,indicating that the relationship between active components and endogenous substances was not a simple linear model,but showed complex pharmacological activity and combined interaction.The results of targeted network pharmacological analysis showed that the ginsenosides absorbed into the blood involved a total of 156 targets.Liver cancer had 2464 targets with strong correlation,and there were 61 common targets,including 27 core targets.In addition,according to the results of pathway enrichment analysis,the main metabolic pathways were involved in cancer metabolic pathways,PI3K-Akt signaling pathway,MAPK signaling pathway,tumor necrosis factor signaling pathway,HIF-1 signaling pathway,insulin resistance,VEGF signaling pathway,apoptosis and so on.It was speculated that the effective parts of Panax ginseng might have a good therapeutic effect on QDLC through the above metabolic pathway.Conclusion:The effective parts of Panax ginseng had obvious therapeutic effect on QDLC,and ginsenosides and polysaccharides had good synergistic effect.In the course of treatment,effective parts of Panax ginseng had great effects on the endogenous substances and gut microbiota in the animal model of QDLC,in which there were many potential biomarkers and gut microbiota with significant differences.Through the treatment of effective parts of Panax ginseng,it regulated the disorder of endogenous substances and gut microbiota caused by pathological model,made it adjust to the healthy state,and played a role in inhibiting tumor growth.In addition,through the analysis of Panax ginseng absorbed into blood and the exploration of the PK-PD during the treatment of QDLC,it showed that the relationship between active components and endogenous substances was not a simple linear model,but showed complex pharmacological activity and combined interaction.Meanwhile,through targeted network pharmacology,the interaction relationship of"active components-action target-disease"was analyzed,which illustrated the material basis and the overall action mechanism. |
PDF Full Text Request