To Explore The Mechanism Of Yinchenhao Decoction In The Treatment Of Liver Cancer Based On Network Pharmacology

Objective: Yinchenhao decoction is a classic prescription of traditional Chinese medicine,which has the effect of clearing heat and dampness,detoxification and treating jaundice.In recent years,clinical and basic studies have shown that Yinchenhao decoction has a certain anticancer effect.On this basis,this study combined network pharmacology and in vitro cell experiments to further explore the effect and mechanism of Yinchenhao decoction on liver cancer,and to provide scientific basis for the clinical application of Yinchenhao decoction.Method:(1)To explore the molecular mechanism of Yinchenhao Decoction on hepatocellular carcinoma through network pharmacology.Firstly,the main active ingredients and related targets of Yinchenhao Decoction were collected by TCM Systematic Pharmacology database and analysis platform(TCMSP),and related targets of hepatocellular carcinoma were obtained through Gene Cards,OMIM and TTD databases.The intersection targets of disease and drug were obtained using Venny platform,and the network diagrams of "drug-active ingredient-target" and "drug-active ingredient-targetdisease" were constructed using Cytoscape3.9.1 software to screen the key components of Yinchenhao Decoction in the treatment of liver cancer.STRING platform was used to construct protein interaction network(PPI)map and analyze the network,so as to obtain the core target of Yinchenhao decoction in the treatment of liver cancer.GO and KEGG enrichment analysis was performed by Metescape platform.Finally,the binding ability of key components to core targets was verified by molecular docking technology,and visualization was performed.(2)The effect of Yinchenhao Decoction on HepG2 cells was investigated by in vitro cell experiments,and the effect of Yinchenhao decoction on HepG2 cell proliferation was investigated by CCK-8 and clonal formation experiments,and half inhibition rate(IC50)was calculated according to the results of CCK-8 experiments.The effect of Yinchenhao Decoction on hepatoma HepG2 cell apoptosis was detected by Hoechst33342 staining assay.The effect of Yinchenhao Decoction on hepatocellular carcinoma HepG2 cell migration was detected by scratch test.The expression levels of corresponding proteins were detected by Western Blot.The mRNA expression level was detected by qPCR.Results:(1)A total of 32 active ingredients and 138 targets of Yinchenhao Decoction in the treatment of liver cancer were obtained.The network diagram of "drug-active ingredienttarget","drug-active ingredient-target-disease" and PPI were constructed,and five key ingredients and core targets were obtained.The top five core targets were AKT1,TP53,VEGFA,TNF,IL6.The top five pathways in KEGG enrichment analysis were PI3K-Akt signaling pathway,cancer signaling pathway,hepatitis B signaling pathway,AGE-RAGE signaling pathway,and chemical cancer-receptor activation signaling pathway.Molecular docking results showed that key components had good binding ability with core targets.(2)The effect of Yinchenhao Decoction on HepG2 cells was investigated in cell experiments.The results of CCK-8 and clonal formation experiments showed that Yinchenhao decoction could significantly inhibit the proliferation of HepG2 cells.The results of Hoechst33342 staining experiments showed that Yinchenhao decoction could induce apoptosis of HepG2 cells.Scratch test results showed that Yinchenhao decoction could inhibit the migration of HepG2 hepatocellular carcinoma cells.Western Blot results showed that after the intervention of Yinchenhao decoction,the protein expression levels of Pi3k,Akt,p-Akt and Bcl-2 in HepG2 cells decreased,while the protein expression level of Bax increased.qPCR results showed that after the intervention of Yinchenhao Decoction,mRNA expression levels of Pi3k,Akt and Bcl-2 in HepG2 cells decreased,while mRNA expression levels of Bax increased.Conclusion:(1)The results of network pharmacological studies showed that Yinchenhao Decoction could intervene on liver cancer by acting on multiple targets and multiple pathways.Quercetin,β-sitosterol,naphthol and other compounds may be the core components of Yinchenhao Decoction in the treatment of liver cancer.(2)Yinchenhao Decoction can inhibit the proliferation of HepG2 hepatoma cells,induce cell apoptosis and reduce the migration ability of hepatoma cells.Pi3k-Akt signaling pathway may be the key pathway for Yinchenhao Decoction to play a role in the treatment of hepatoma.