Based On Network Pharmacology And Molecular Docking To Study The Mechanism Of Shisiwei Wendan Decoction In The Intervention Of Lung Adenocarcinoma

Objective To study the pathway and target of Shisiwei Wendan Tang(SWT)in the treatment of lung adenocarcinoma(LUAD)based on network pharmacology and molecular docking.Cytological and animal experiments proved that SWT regulated autophagy,apoptosis,proliferation and migration of cells through the HSP90AA1/AKT/mTOR pathway,and then intervened in the occurrence and progression of lung adenocarcinoma.Finally,the main mechanism of action of SWT intervention in LUAD was revealed.Methods 1.Use the drug online database and literature to obtain SWT drug active ingredients and targets;Using disease databases to predict disease targets for lung adenocarcinoma;Draw Venn diagrams and Disease-Drug-active ingredient-Key target regulatory networks using R.Protein-Protein Interaction Networks(PPI)were analyzed using STRING database.The Biological Processes(GO)and the Kyoto Encyclopedia of Genes and Genomes(KEGG)were analyzed based on the Metascape platform;their genomes were analyzed jointly;Molecular docking was studied using Autodock vina.Finally,Cytoscape was utilized to perform visual analysis on the above data results.2.Prepare SWT-containing serum to interfere with A549 cells,and set SWT-containing serum low,medium and high dose groups,negative serum groups and control group.LC3 Turnover method was used to detect the protein expressions of LC3 II and LC3 I before and after intervention to evaluate the effect of drug-containing serum on autophagy level.The cell proliferation was detected by CCK-8 method after the intervention of drug-containing serum.Apoptosis was detected by flow cytometry to evaluate the effect of drug-containing serum on apoptosis.The effect of drug-containing serum on cell invasion was evaluated by cell scratch test.The protein expression levels of HSP90AAA1,P-AKT,AKT,P-mTOR and mTOR in A549 cells were detected by WB to determine whether HSP90AA1/AKT/mTOR pathway was the target and pathway for SWT to intervene LUAD in vitro.3.A549 lung adenocarcinoma cells were used to establish a subcutaneous transplantation tumor lung cancer model in nude mice,which was divided into model group,cisplatin group,SWT low-dose,medium-dose and high-dose groups for 30 days.The weight changes of mice were observed and the mass of subcutaneous tumor was measured.HE staining,TUNEL staining and Ki67 immunohistochemical staining were used to evaluate the pathological morphology,cell apoptosis and cell proliferation.Autophagy was evaluated by WB detection of LC3Ⅱ/Ⅰ.The protein expression levels of HSP90AAA1,P-AKT,AKT,P-mTOR and mTOR in lung cancer tissues were detected by WB to determine whether the HSP90AA1/AKT/mTOR pathway is the target and pathway for SWT to interfere with LUAD in vivo.Results 1.Network pharmacology and molecular docking studies have shown that SWT regulates the expression of HSP90AA1,AKT1,MAPK1,MAPK3 and TP53 through active ingredients such as tanetin,dihydrotanetin,naringin,carotenin,quercetin and kaempferol.Then,it interferes with tumor necrosis factor signaling pathway,chemical carcinogenic receptor activation pathway,Th17 cell differentiation signaling pathway and other pathways involved in apoptosis,autophagy,negative regulation of cell proliferation,homeostasis of multicellular organism,response to oxidative stress,response to lipopolysaccharide and other aspects to play a role in LUAD treatment.It reflects the multi-target,multi-component,multi-pathway and multi-dimensional network pharmacological characteristics of SWT.2.Cell experiments showed that low,medium and high doses of SWT drug-containing serum could all up-regulate the fine apoptosis and autophagy levels of A549,and inhibit cell proliferation and invasion.3.Animal experimental studies have found that intervention with high,medium and low concentrations of SWT can delay the progression of LUAD and reduce tumor weight.After the intervention of Shishiwei Wendan Decoction,the expression of autophagy marker protein LC3-II in tumor tissues was significantly increased,while the expression of LC3-I protein was decreased(P < 0.01),indicating that autophagy level was significantly increased.Further histopathological tests showed that SWT could promote cell apoptosis and inhibit cell proliferation,and play its role in inhibiting the progression of LUAD.4.In both cell and animal experiments,SWT can inhibit the expression of HSP90AA1 protein,inhibit the phosphorylation of AKT and mTOR,promote cell apoptosis and autophagy,and reduce cell proliferation and migration ability.These results suggest that SWT inhibits the development of LUAD by inhibiting the HSP90AA1/AKT/mTOR signaling pathway.Conclusion 1.The results confirmed that SWT inhibited the expression of HSP90AA1 in A549 cells,and then inhibited the phosphorylation of AKT and mTOR,so as to promote the autophagy and apoptosis of A549 cells,inhibit the proliferation and migration of lung adenocarcinoma cells,and ultimately delay the progression of lung adenocarcinoma tumors.2.SWT can inhibit the proliferation of LUAD,up-regulate the autophagy level of LUAD and promote the apoptosis of LUAD by inhibiting the activity of HSP90AA1/Akt/mTOR signaling pathway.Thus,it plays an antitumor role in vivo and in vitro.