| Blumea balsamifera belongs to Blumea,Compositae.It can be used as a medicine in whole grass or on the ground.It is bitter and cold,and enters the spleen and lung channels.According to the literatures,the main ingredients of B.balsamifera flavonoids,volatile oils and terpenes are,and also contain a small amount of coumarin,sterols,organic acids,amino acids and other components,which have pharmacological effects such as antitumor,antioxidant,analgesic and antibacterial.The purpose of this study was to separate a series of compounds from the n-butanol extract of B.balsamifera,and study the anti-NSCLC activity of the n-butanol extract and compounds.The network pharmacology method was used to preliminary explore the correlation between the chemical constituents acting on non-small cell lung cancer.These laid the foundation for analyzing the pharmacodynamic substance basis and potential mechanism of B.balsamifera again NSCLC.The research content is mainly divided into three parts as follows:1.10 compounds were separated from the n-butanol extraction of B.balsamifera with using ethanol reflux extraction,silica gel,macroporous resin,gel and other column chromatography and recrystallization and other methods for separation.According to the physical and chemical properties and spectral data,and they were identified as luteolin-7-O-?-D-glucuronide methyl ester(1),luteolin(2),luteolin-7-O-?-D-glucuronide(3),luteolin-7-O-?-D-glucoside(4),apigenin(5),ethyl caffeate(6),luteolin-7-O-?-D-glucosyl(4?1)-?-L-rhamnopyranoside(7)and xanthoxylin(8).Compounds 1,3 and 4 were obtainted from this plant for the first time,and Compound 7 is novel flavonoid glycoside.The special reagent inspection method was used to characterize the type of compound in the n-butanol extract.The results indicate that the extract mainly contains flavonoids and their glycoside,tannins or phenols.The total flavonoid content of n-butanol extraction was determined with chemical reagent[NaNO2-Al(NO3)3-NaOH].The results showed that the total flavonoid content of the n-butanol extraction was 42.17%(mg/mg),which was higher than the total flavonoid content of the extract(17.43%,mg/mg).2.The CCK-8 method was used to investigate the proliferation inhibition and effect of n-butanol extract and its compounds on A549,NCI-H1299 cells.The results showed that the n-butanol extract had a certain inhibitory effect on the proliferation of these two cells,and the inhibitory effect was dependent on the concentration of the drug,the IC50 of A549 and NCI-H1299 cells were 333.10?g/mL and 422.00 ?g/mL.Compounds 2 and 5 showed better cytotoxic activity than compounds 1,4 and 7,with IC50 of 41.2 ?M and 63.85 ?M,respectively.When the drug concentration was 100 ?M,the proliferation inhibition rates of Compounds 1,4 and 7 were 20.71%,14.65%and 15.93%,respectively,and the IC50 was greater than 100 ?M.3.The compounds were screened for activity according to rules of Lipinski and Veber.The target prediction of the compounds was performed through the TCMSP database and the SwissTarget Prediction platform.The Gene Ontology and KEGG pathway analysis of the predicted target is through Cytoscape 3.7.2.Finally,target verification was performed by Autodock Vina.The results showed that:29 active ingredients were screened from B.balsamifera,and 84 targets were predicted to act on NSCLC.Among the entries,86 entries are related to biological processes,6 entries are related to cellular components,and 16 entries are related to molecular functions.It is related to 40 KEGG pathways,involving tumor-related signaling pathways and metabolic signaling pathways.The core targets PIK3R1,EGFR,SRC,and AKT1 not only contribute greatly to the PPI network,but also participate in multiple KEGG pathways.EGFR,AKT1,and SRC are docked with their corresponding compounds(F23,F27,F30,and F32).The binding energy between them is less than-5 kcal/moL,indicating that the compound as a ligand molecule can spontaneously bind to the receptor protein. |
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