Study On Pharmacodynamics Mechanism Of Diwu Anti-liver Cancer Based On Metabonomics And Transcriptomics

Objective: To evaluate the antitumor efficacy of Chinese medicine Diwu in vivo by establishing a subcutaneous xenograft tumor model of H22 hepatocellular carcinoma cells in nude mice.Analytical experiments were performed to find differential metabolites and differentially expressed genes,to analyze the relevant metabolic pathways and signal pathways,and to explain the biological connotation of the mechanism of Diwu treatment of liver cancer from the perspective of metabolomics and transcriptomics.Methods: Twenty-four BALB/C-nu male nude mice were randomly divided into four groups: model group(saponin 0 mg/kg,physiological saline),low-dose group(saponin 5 mg/kg),medium-dose group(saponin 10 mg/kg)and high-dose group(saponin 15 mg/kg),6 rats in each group.The tumor tissue was washed with saline after 24 h and weighed to calculate tumor inhibition rate.The tumor tissue growth curve was plotted.Metabonomics analysis and RNA-seq were carried out in model group and high dose group Analysis.Results:(1)Pharmacodynamic results: The results showed that different concentrations of total diurin could significantly inhibit tumor growth in nude mice,and the behavior and mental condition of nude mice in the drug groupwere good,indicating that there was no obvious side effect in gavage administration of traditional Chinese medicine.The results showed that the tumor growth curve showed that the tumor growth curve showed that the tumor tissue growth could be effectively inhibited,among which the high-dose group had the most significant inhibitory effect;the tumor tissue weight of each group was significantly reduced compared with the model control group all of these differences were significant(p<0.01).And the tumor inhibition rates of the high,middle and low dose groups were 66.54%,55.73% and53.36%,respectively.HE staining sections showed that all the dose groups of Diwu could promote tumor cell death in different degrees.(2)Metabolomics results: A total of 71 peaks were detected in the GC-MS experiment,of which 30 small-molecule metabolites were mostly identified as amino acids and fatty acids,and a few sugars,esters,and alcohols.From the trend of content change,compared with the model group,after the intervention of diurtriterpenoid saponin,dihydrocarvitol,2-hydroxyvaleric acid,sodium4-hydroxybutyrate,aspartic acid,dehydrated inositol,Isopropanolamine,erythrose,gluconolactone,glutamic acid,glutaric acid,histidine methyl palmitate,N-acetyl-L-ornithine,pantothenic acid,pentadecanoic acid,sulfuric acid,17 metabolites including tetrahydrocorticosterone were reduced.Preliminary identification of 9 differential metabolites in tumor tissues,including 2-hydroxyvaleric acid,aspartic acid,gluconolactone,glutamic acid,glutaric acid,guanidinoacetic acid,ribitol,and histidine 2N-?-acetyl-L-ornithine.Differential metabolites are mainly enriched in Histidine metabolism,Nitrogen metabolism,Arginine and proline metabolism,Beta-alanine metabolism,Pentose phosphate pathway,Alanine,aspartate and glutamate metabolism.Histidine metabolism is the most closely related to differential metabolites.The anti-hepatocellular carcinoma mechanism may significantly reduce the expression of L-histidine and aspartic acid in thehistidine metabolism pathway,and inhibit tumor cells.Signal transduction and proliferation,play an antitumor effect.(3)Transcriptomics results: The statistical results of differentially expressed genes between different groups showed that after drug administration intervention,a total of 202 genes had significant changes in expression levels,among which 5 genes significantly increased the expression level and 197 significantly decreased(p <0.01).These differential genes are enriched in Hematopoietic cell lineage,Cytokine-cytokine receptor interaction,MAPK signaling pathway,PI3K-Akt signaling pathway,PPAR signaling pathway,Chemokine signaling pathway and other pathways,which affect the proliferation and immunity of liver cancer cells.Among them,there are many differential genes enriched under the PI3K-Akt signaling pathway,and there are significant differences.Differential expression of Pik3 cg,Akt,Mtor,and Pten in related signal pathways was detected by qRT-PCR.The results showed that these four genes were all expressed in tumor tissues.Compared with the model group,diwu treatment significantly increased the expression level of Pten gene and down-regulated the expression levels of Pik3 cg,Akt1,and Mtor genes.Conclusions: 1.Diwu can effectively inhibit the growth of tumor tissues,and the inhibitory effect of the high-dose group(Diosgenin 15 mg / kg)is the most significant.2.Histidine metabolism may play a role in inhibiting the proliferation of liver cancer cells by diwu.Inhibition of PI3 K / AKT mTOR signaling pathway may play a role in the inhibition of hepatoma cell proliferation bydiwu.