| Selaginella tamariscina(Beauv.)Spring is a dried whole plant of Selaginellaceae(Selaginellaceae)Selaginella.The 2020 Chinese Pharmacopoeia records that it has the effect of promoting blood circulation and regulating menstruation,and is used for amenorrhea and dysmenorrhea.,Zheng scars,lump,fall and fall injury.Selaginella charcoal removes blood stasis and stops bleeding,and is used for vomiting blood,metrorrhagia,blood in the stool,and prolapse of the anus.It was listed as top grade in "Shen Nong’s Materia Medica".Modern pharmacological research shows that Selaginella has anti-tumor,hypoglycemic,antibacterial,anti-inflammatory and antioxidant effects.This thesis researches the chemical composition of Selaginella by means of traditional natural product analysis techniques and various chromatographic methods.Preliminary predictions of the anti-tumor active ingredients,targets and pathways of Selaginella vulgaris were carried out using the method of network pharmacology.The in vitro anti-tumor activity of the components of Selaginella was investigated to lay the foundation for the research on the material basis of the anti-tumor efficacy of Selaginella.The research content is mainly divided into three parts,as follows:1.Through flash extraction method,solvent extraction method,macroporous adsorption resin column chromatography,silica gel column chromatography,polyamide column chromatography,ODS column chromatography and preparative high performance liquid methods,10 were separated from the ethyl acetate part of Selaginella The compounds were identified by physicochemical properties and spectroscopy methods.They are selaginellin(1),selaginellin M(2),selaginellin A(3),diselaginellin A(4),diselaginellin B(5),and selaginellin E(6),kaempferol-3-O-α-L-rhamnoside(7),apigenin-7-O-β-D-glucoside(8),apigenin-4’-O-βD-glucoside(9),kaempferol-3-O-(6"-O-Acetyl)-β-D-glucopyranoside(10).Among them,compounds 8,9,and 10 were isolated from Selaginella for the first time.2.Using network pharmacology methods to preliminarily explore the anti-tumor active ingredients of Selaginella and its mechanism of action,use TCMSP,SymMap,ETCM database and related literature to screen the active ingredients of Selaginella,use TCMSP,Pubmed,Swiss Target Prediction database to compare The potential targets of the compounds were predicted,and the DAVID6.7 database was used to perform GO annotation and KEGG pathway analysis on the predicted core targets.The results showed that 12 main anti-tumor active ingredients were screened from Selaginella,including 4-HydroxyMephenytion,Apigenin,Azaleatin,Caffeic acid,Dihydroresveratrol,Farrerol,Fastigilin B,Gossypetin,Kaempferol,Selaginellin E,Hinokiflavone,Ferulaic acid.There are 11 predicted core antitumor targets of Selaginella,namely AKT1,EGFR,ESR1,SRC,PTGS2,PIK3CA,MMP9,AR,CDK4,KDR and MMP2.The KEGG-enriched pathways screened 20 signaling pathways,namely Pathways in cancer,Endocrine resistance,Proteoglycans in cancer,Estrogen signaling pathway,Relaxin signaling pathway,Bladder cancer,Prostate cancer,and Breast cancer.3.The MTT method was used to investigate the cell proliferation inhibitory effects of Selaginella elata extract on human breast cancer cells MCF-7,human gastric cancer cells SGC7901,and human liver cancer cells HepG-2.The results showed that Selaginella extracts have a certain inhibitory effect on the three tumor cells,showing a significant dose-effect relationship.Among them,Selaginella has a significant inhibitory effect on the human breast cancer cell MCF-7.The MTT method was used to investigate the inhibitory effects of Selaginella petroleum ether,chloroform,ethyl acetate,n-butanol and water extract on the proliferation of human breast cancer cell MCF-7.The experimental results show that each site has different degrees of inhibition,and the ethyl acetate extraction site has the best anti-tumor activity.MTT method and flow cytometry were used to investigate the compounds selaginellin(1),selaginellin A(3),selaginellin E(6),amentoflavone(11),selariscinin D(12),selaginellin T(13),apigenin(14)Inhibitory activity on human breast cancer cell MCF-7.The results show that each compound has a certain activity and is dose-dependent within a certain concentration.Among them,compound 1 shows a good inhibitory effect.Flow cytometry is used to determine whether compound 1 can induce MCF-7 cell apoptosis.analysis.The results showed that as the concentration of compound 1 increased,the rate of apoptosis also increased,suggesting that the induction of apoptosis may be an important mechanism for Selaginella to inhibit MCF-7 cells.In summary,10 compounds were separated from the ethyl acetate part of Selaginella for this project,including 5 acetylenic phenolic compounds and 5 flavonoids,and 3 of them were isolated from Selaginella for the first time.In vitro anti-tumor studies found that compound 1 inhibited the proliferation of MCF-7 cells more significantly,followed by compound 6.It also verified that selaginellin E predicted by network pharmacology does have anti-breast cancer activity.The research of this subject has laid the foundation for Selaginella antitumor. |
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