To Explore the Mechanism of Huangqin Decoction(黄芩汤)in the Treatment of Liver Cancer Based on Network Pharmacology,Molecular Docking Technology and In Vitro Experimental Verification
Objective:To study the material basis and potential mechanism of Huangqin decoction in the treatment of hepatocellular carcinoma by network pharmacology and molecular docking,and to verify it by in vitro experiments.Methods:The active components and targets of Huangqin(Scutellaria baicalensis),Shaoyao(Paeonia lactiflora),Dazao(jujube)and Gancao(licorice)in Huangqin decoction were searched through the traditional Chinese medicine system pharmacological platform(TCMSP)database.The active components and targets were screened under the conditions of oral bioavailability(OB)≥30％and DL≥0.18,and the target was predicted by Uniprot database.With"liver cancer"as the key word,the disease target of liver cancer was obtained by GeneCards,OMIM and DrugBank database.The drug action target and the disease target were intersected by Wayne diagram,and the PPI protein interaction network map was constructed by String database.The exported tsv data file was imported into Cytoscape 3.8.2 software to further screen the core target.The disease and drug targets were imported into Cytoscape software,to construct the network topology map of the interaction between drug PPI and disease PPI protein,and the core targets were selected.The"drug-active ingredient-target"network diagram was constructed by Cytoscape software.The DAVID database was used for gene ontology(GO)enrichment analysis and Kyoto Encyclopedia of Gene and Genome(KEGG)pathway enrichment analysis.Python script and AutoDuck Vina 1.2.0 software were used to dock the core components of Huangqin decoction with key targets,and the binding force was calculated.HepG2 proliferation was detected by CCK-8.Apoptosis was detected by TUNEL staining.mRNA level of core target was detected by q-PCR,and predictive pathway was verified by Western blotting experiment.Results:Totally 160 active components were selected from Huangqin decoction,and there were 238 corresponding active targets.The number of disease corresponding targets was 1 474 after screening and removing duplicates,and the intersection number was 120.The enrichment analysis of KEGG and GO showed that there were PI3K-AKT signal pathway,cancer signal pathway,hepatitis B signal pathway,AGE-RAGE signal pathway,hepatitis C signal pathway,IL-17 signal pathway,TNF signal pathway and so on.BP is mainly concentrated in the positive regulation of gene expression,the negative regulation of gene expression,the positive regulation of RNA polymerase Ⅱ promoter transcription,the negative regulation of apoptosis,the positive regulation of apoptosis,the response of cells to tumor necrosis factor and so on.The results of molecular docking showed that the molecular docking binding strength of quercetin,kaempferol,[3-sitosterol,wogonin and baicalein with TP53,AKT1,CASP3,JUN and VEGFA was stable.Cell experiments showed that the serum containing Huangqin decoction could inhibit the proliferation and apoptosis of HepG2 cells,up-regulate the expression of TP53 mRNA,CASP3 mRNA and PTEN mRNA,down-regulate the expression of AKT1 mRNA,and decrease the protein expression of p-PI3K and p-AKT.Conclusion:The main therapeutic effect of Huangqin decoction on hepatocellular carcinoma is to enhance the expression of tumor suppressor gene PTEN and down-regulate the expression of p-PI3K and p-AKT,so as to inhibit PI3K/AKT signal pathway and induce apoptosis of hepatoma cells to weaken the proliferation of HepG2 cells.
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维普
