| Objective: Separation of the n-butanol extract of Scutellaria barbata, obtained a series of fractions and compounds, and study anti endometrial cancer cell activity in vitro and anti-endometrial cancer cell activity, screened with anti endometrial cancer cell active component or components, and to explore anti-endometrial cancer activity and content of chemical components between correlation and lay the foundation for the next research of Scutellaria barbata anti-endometrial cancer cell mechanism.Method: By reflux and solvent extraction, vacuum drying method for preparingn-butanol; By using macroporous resin, silica gel column chromatography and Sephadex LH-20 and combined with HPLC separation guide n-butanol fraction; NMR techniques of Herba Scutellariae barbatae is butanol fraction isolated from the monomer of analysis, to determine the structure of the compound; MTT method of Herba Scutellariae barbatae is butanol fractions and monomer activity screening; plate scratch assay and Transwell culture method for the detection of endometrial cancell migration ability; PI staining, the cell cycle was detected by flow cytometry, by HPLC-MS of Herba Scutellariae barbatae n-butanol activity group points to study the chemical constituents.Results: Scutellaria barbata n-butanol headquarters were preliminarily isolated five fractions, MTT activity screening, found Fr.2 group, Fr.3 group, Fr.4 group activity was better than others. The for further separation were isolated and identified eight compounds,including four flavones and two flavonoid glycosides, 1 sterol, a benzene formyl derivatives. The four flavonoids: 5, 6, 7- trihydroxy- 4′- methoxy two hydrogen flavonoids(compound 1), 4′- hydroxy wogonin(compound 2), celery(compound 6),luteolin(compound 7); two flavonoid glycosides: Different Alpine Scutellaria element8-O- glucuronide(compound 3), scutellarin(compound 4); 1 sterol: beta sitosterol(compound 8); 1 benzoyl derivatives: hydroxy benzaldehyde(compound 5). The compounds 1 and 3 were the first isolated from Scutellaria barbata. And the MTT activity of the monomer compounds was detected. It was found that the effect of the anti endometrial cancer was stronger than that of other compounds.and its IC50 value was135.603 μg·mL-1. The MTT activity was screened by the Fr.4.2 activity of the normal butyl alcohol, and it was found that the IC50 value was 37.435 μg·mL-1, and the activity was the strongest. The activity of Fr.4.2 were separated by 4′, 5, 7- three hydroxyflavone(apigenin). Compared with the control group, the Ishikawa cells by 25, 80, 100μg·mL-1Fr.4.2 group for testing medicine treatment of G2 phasecells increased significantly after 24 h; using HPLC-MS of Herba Scutellariae barbatae normal butanol activity group are analyzed. Through references that are relevant to infer the 13 possible compounds.Conclusion: Activity was detected by MTT activity of the isolated compounds from the normal butanol, and the inhibitory effect of Baicalin on Ishikawa cells was stronger than that of other single compounds. Is the butanol active component can significantly influence the Ishikawa cell cycle progression, cell cycle arrest in the G2 phase, suggesting that the anticancer mechanism may be through the influence of G2 protein synthesis,prevents cells from entering a division cycle, thereby inhibiting the proliferation of endometrial cancer cells, achieve the anti-cancer effect. The activity of HPLC-MS group of Scutellaria barbata n-butanol are analyzed that the flavonoids of Scutellaria barbata as the main active component of anti endometrial cancer, laid the foundation for the study of the mechanism for the next step in the research of anti endometrial cancer scutellariabarbata. |
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