Study On The Material Basis And Mechanism Of Action Of Poria Cocos Against Nasopharyngeal Carcinoma Based On Network Pharmacology And Molecular Docking Technolog

Objective: To explore the potential material basis and action mechanism of poria cocos against nasopharyngeal carcinoma by using network pharmacology,molecular docking technology and cell experiment.Methods: 1.The databases of TCMSP and BATMAN-TCM were used to retrieve the potential active chemical components and targets of Poria cocos.The Gencards and OMIM databases were used to establish a nasopharyngeal carcinoma target database and map it to Poria cocos target,and the Poria cocos–nasopharyngeal carcinoma intersection target was analyzed for PPI using STRING database to screen potential core targets.The DAVID database was used for GO and KEGG enrichment analysis to predict the involved biological processes and metabolic pathways.2.The molecular docking technology was used to verify the binding activities of poria cocos acid in Poria cocos active components and TP53,ESR1,PRG and ANXA1 targets in PPI analysis results.3.CCK-8 method and RTCA method were used to monitor the effect of pachymic acid on the proliferation of nasopharyngeal carcinoma CNE2 cells.Fluorescence double-staining flow cytometry and Hochest33342 were used to detect the effect of pachymic acid on the apoptosis of CNE2 cells.Western blot was used to detect the expressions of p53,ESR1 and PI3K/Akt signaling pathway-related proteins.Results: 1.In this study,eight active chemical components represented by Poria cocos acid,Poria cocos new acid B,and hederagenin were screened out,and 18 core targets represented by TP53,INS,ESR1,and PTGS2 were identified.GO analysis mainly involves biological processes such as cytokine activity,immune inflammatory system,and tumor cell proliferation,apoptosis,and migration.KEGG analysis is mainly enriched in thyroid hormone signaling pathway,estrogen signaling pathway,PI3K/Akt signaling pathway,etc.2.The molecular docking results showed that the free energy of binding ofpachymic acid to TP53,ESR1,PRG,and ANXA1 was less than 0 kcal/mol,indicating that pachymic acid had good binding to the targets selected by PPI.3.CCK-8 results showed that,compared with the control group,Poria cocos acid had a significant inhibitory effect on the proliferation of nasopharyngeal carcinoma CNE2 cells(P<0.05 or P<0.01).Fluorescence double-staining flow cytometry and Hochest33342 staining showed that Poria cocos acid could induce apoptosis of CNE2 cells(P < 0.05 or P < 0.01).Western blot results showed that compared with the control group,pachymic acid could reduce the expression levels of p53,PGR and ESR1 proteins(P < 0.05 or P < 0.01),and reduce the expression levels of PI3 K and p-Akt,the key proteins of PI3K/Akt signaling pathway in CNE2 cells(P < 0.05 or P < 0.01).However,there was no significant difference in the expression of Akt(P > 0.05).After pachymic acid was added with PI3K/Akt signaling pathway activator SC79,RTCA and fluorescence double-staining flow cytometry showed that the proliferation inhibition and apoptosis-inducing effect of pachymic acid on CNE2 cells were weakened.Conclusion: This study preliminarily predictes the effective chemical components,targets,possible biological processes and signaling pathways of Poria cocos in the treatment of nasopharyngeal carcinoma.Pachymic acid,a representative active chemical component of Poria cocos,may inhibit the proliferation of nasopharyngeal carcinoma cells and induce apoptosis by inhibiting p53,ESR1 and PI3K/Akt signaling pathways.