Network-based Pharmacological Study Of The Mechanism Of Ruan Gan Li Dan Tang For The Treatment Of Liver Cancer

Objective: Traditional Chinese medicine has a long history and unique and effective experience in the treatment of liver cancer.It is widely applied to patients with all stages of liver cancer,and is effective in promoting blood circulation,dispersing nodules and relieving pain.After clinical use,it was found that Ruan Gan Li Dan Tang has the effects of reducing the rate of postoperative recurrence and metastasis,alleviating the side effects of interventional and radiotherapy,improving the quality of life and prolonging the survival period of patients with hepatocellular carcinoma.This study combines network pharmacology with cell experiments to explore the mechanism of action of Ruangan Lidan Tang on liver cancer,providing scientific basis for clinical application.Methods:(1)We collected the drug targets and related components of HCC in the pharmacological database and analysis platform of TCM system,and searched and screened the genes of HCC disease targets in Gene Cards and OMIM database.The drug-disease intersection targets were analyzed by constructing protein interaction network,"drug-disease" target network,KEGG pathway enrichment analysis and GO function enrichment analysis,molecular docking,etc.,to predict the possible targets and main active ingredients of Hep G2 for hepatocellular carcinoma.(2)After obtaining Hep G2 hepatocellular carcinoma cell line,cell culture was performed,and the lyophilized powder of Ruan Gan Li Dan Tang was divided into low-dose group,medium-dose group and high-dose group,and a blank control group was set up to add Hep G2 hepatocellular carcinoma cell line,and cell proliferation was observed by CCK8 method,cell invasion ability was observed by Transwell assay,migration ability of hepatocellular carcinoma cells was detected by scratch assay,and protein blotting assay The expression of pathway-related proteins was detected by CCK8 assay,Transwell assay for cell invasion,scratch assay for migration,and protein imprinting assay.Results: Eleven drugs,including Chaihu,Huangqin,Yanhusuo,Turmeric,Tianjihuang,Chupencao,Yinchen,Prunella,Red Ginseng,Pinellia ternata,and Glycyrrhiza,were searched through the TCMSP platform database.Under the conditions of OB ≥ 30% and DL ≥ 0.18,216 potential compounds and 306 drug targets of Ruangan Lidan Tang were obtained.Intersecting drug targets with disease targets yields 25 common targets.At the same time,25 common targets were uploaded to the DAVID online website for GO enrichment analysis and KEGG pathway enrichment analysis.GO functional enrichment analysis showed that the targets of Ruangan Lidan Tang participated in the biological processes,cellular components,and molecular functions,with the highest ranking being positive regulation of gene expression,cellular response to drug,and receptor complex,respectively.The top five pathways are proteoglycans in cancer,tumor pathways,micro RNAs in cancer,PI3K-Akt signaling pathway,and HIF1α Signal pathway.According to the "Disease Drug Target" network,the top 5 potential core targets for treating liver cancer are TP53,MDM2,STAT3,HIF1 a,AKT1,CCND1.We used molecular docking technology to analyze the binding energy of the five main active ingredients and the five main potential targets in Ruangan Lidan Decoction.The docking scores were all less than-6,indicating that the main active ingredients of Ruangan Lidan Decoction had certain binding activity with the potential targets for treating liver cancer.It can be speculated that Ruangan Lidan Decoction may act on liver cancer through the above pathways and targets.In the cell experiment,the CCK8 experiment showed that after the intervention of Ruangan Lidan Tang,the proliferation of liver cancer cells was inhibited in a dose-dependent manner;Immunoblotting experiments showed that after intervention with Ruangan Lidan Tang,the expression of PI3 K,Akt,p-Akt,and Bcl-2 proteins in liver cancer cells was inhibited,while the expression of Bax protein was increased.We speculate that Ruangan Lidan Tang may affect Hep G2 liver cancer cells through the PI3K/Akt signaling pathway.Conclusion:This study predicts the mechanism of Ruangan Lidan Tang in treating liver cancer through network pharmacology,and the results indicate that the target of Ruangan Lidan Tang is closely related to liver cancer;It involves many signaling pathways that regulate the occurrence and development of liver cancer,such as the PI3K/Akt signaling pathway.The CCK8 experiment verified that Ruangan Lidan Tang inhibits the proliferation of liver cancer cells.Immunoblotting experiments detected that the expression of key proteins in the PI3K/Akt signaling pathway was inhibited.We speculate that Ruangan Lidan Tang can regulate Hep G2 liver cancer cells through the PI3K/Akt signaling pathway.This study provides a scientific basis for in-depth exploration of the molecular mechanism of Ruangan Lidan Tang in the treatment of liver cancer.